Specific Lactate Dehydrogenase Research Articles

Serological profile of voluntary blood donors in a blood bank of tertiary health care centre- An institution-based study

Background: The safe transfusion of blood facilities is an essential component of an efficient, excellent healthcare sector. Unsafe blood transfusions can be a cause of transfusion-transmitted infections and can be lethal. The present study was conducted to find out the seroprevalence of HBsAg, HCV, HIV, malarial parasites, and syphilis in voluntary blood donors in a tertiary care center. Materials and methods: This prospective study was conducted at the blood bank of the Shri Mahant Indiresh Hospital, Dehradun. 16413 samples from all blood donations were screened for HBsAg, HCV, HIV 1 and 2, malaria, and syphilis. Samples were collected in vacutainers at the time of blood donation and screened for HIV 1and2, HBsAg, and HCV using the fourth-generation enzyme-linked immunosorbent assay technique. Syphilis was tested by Treponema antibodies using the VITROS Syphilis TPA Reagent Pack. Malaria was tested by pan malaria card, monoclonal anti-pan specific parasite lactate dehydrogenase antibody conjugated to colloidal gold, and another monoclonal anti-pan specific parasite lactate dehydrogenase antibody immobilized on a nitrocellulose strip in a thin line. Results: Out of 16413 blood donors 342(2.1%) were found to be seropositive and 16071 (97.9%) were found to be seronegative. The total number of donors, who were found positive for HBsAg/Anti HCV was 1 (0.3%). Out of the seropositive cases, HBsAg, HCV, HIV, and Syphilis infections constituted 28.9%, 27.5, 14.6, and 28.7% cases respectively. Conclusions: There is a need for the development and implementation of stringent donor selection and sensitive screening tests to reduce the risk of acquiring transfusion-transmitted illnesses.

Castration-resistant prostate cancer monitoring by cell-free circulating biomarkers.

Prostate cancer is the second leading cause of male cancer-related deaths in Western countries, which is predominantly attributed to the metastatic castration-resistant stage of the disease (CRPC). There is an urgent need for better prognostic and predictive biomarkers, particularly for androgen receptor targeted agents and taxanes. We have searched the PubMed database for original articles and meta-analyses providing information on blood-based markers for castration-resistant prostate cancer monitoring, risk group stratification and prediction of therapy response. The molecular markers are discussed along with the standard clinical parameters, such as prostate specific antigen, lactate dehydrogenase or C-reactive protein. Androgen receptor (AR) alterations are commonly associated with progression to CRPC. These include amplification of AR and its enhancer, point mutations and splice variants. Among DNA methylations, a novel 5-hydroxymethylcytosine activation marker of TOP2A and EZH2 has been identified for the aggressive disease. miR-375 is currently the most promising candidate among non-coding RNAs and sphingolipid analysis has recently emerged as a novel approach. The promising biomarkers have the potential to improve the care of metastatic prostate cancer patients, however, they need further validation for routine implementation.

A New Role for Lactate in Breast Cancer

Lactate dehydrogenase (LDH) is an enzyme that plays a key role in cellular metabolism by converting pyruvate to lactate during anaerobic glycolysis. LDH has been implicated in various cancer types, including breast cancer, where increased LDH activity is associated with tumor growth, invasion, and resistance to therapy. LDH inhibitors are being investigated as potential therapeutic agents in cancer treatment, including breast cancer. Khajah and colleagues investigated the role of LDH inhibitors in both estrogen receptor (ER)+ and ER‑ breast cancer cell lines and in normal breast epithelial cells. A significant reduction in extracellular lactate level, cell proliferation, motility, and invasion was observed after treatment with LDH inhibitors. Subsequently, changes in the levels of vimentin, E‑cadherin, p38 MAPK, ERK1/2, and AKT were also observed. Cancer cell proliferation was preferentially blocked compared with normal epithelial cell inhibition by LDH inhibitors such as quercetin and lonidamine. These results further highlight the role of lactate in breast cancer progression and indicate the practical use of various commercially available LDH inhibitors as promising therapeutic agents to oppose the processes leading to cancer progression. While LDH inhibitors show promise, challenges include achieving selectivity for cancer cells and minimizing toxicity to normal tissues. The development of specific LDH inhibitors with favorable pharmacokinetic properties is an ongoing area of research. Mol Med Rep. 2024 Jan;29(1):12. doi: 10.3892/mmr.2023.1313.

The effect of electrical stimulation of skeletal muscle on cardioprotection and on muscle-derived myokine levels in rats: A pilot study.

Electrical muscle stimulation (EMS) is a widely used method in sports and rehabilitation therapies to simulate physical exercise. EMS treatment via skeletal muscle activity improves the cardiovascular functions and the overall physical condition of the patients. However, the cardioprotective effect of EMS has not been proven so far, therefore, the aim of this study was to investigate the potential cardiac conditioning effect of EMS in an animal model. Low-frequency 35-min EMS was applied to the gastrocnemius muscle of male Wistar rats for three consecutive days. Their isolated hearts were then subjected to 30 min global ischemia and 120 min reperfusion. At the end of reperfusion cardiac specific creatine kinase (CK-MB) and lactate dehydrogenase (LDH) enzyme release and myocardial infarct size were determined. Additionally, skeletal muscle-driven myokine expression and release were also assessed. Phosphorylation of cardioprotective signaling pathway members AKT, ERK1/2, and STAT3 proteins were also measured. EMS significantly attenuated cardiac LDH and CK-MB enzyme activities in the coronary effluents at the end of the ex vivo reperfusion. EMS treatment considerably altered the myokine content of the stimulated gastrocnemius muscle without altering circulating myokine levels in the serum. Additionally, phosphorylation of cardiac AKT, ERK1/2, and STAT3 was not significantly different in the two groups. Despite the lack of significant infarct size reduction, the EMS treatment seems to influence the course of cellular damage due to ischemia/reperfusion and favorably modifies skeletal muscle myokine expressions. Our results suggest that EMS may have a protective effect on the myocardium, however, further optimization is required.

Construction and validation of a clinical predictive nomogram for intraductal carcinoma of the prostate based on Chinese multicenter clinical data.

Intraductal carcinoma of the prostate (IDC-P) is a special pathological type of prostate cancer that is highly aggressive with poor prognostic outcomes. To establish an effective predictive model for predicting IDC-P. Data for 3185 patients diagnosed with prostate cancer at three medical centers in China from October 2012 to April 2022 were retrospectively analyzed. One cohort (G cohort) consisting of 2384 patients from Zhejiang Provincial People's Hospital was selected for construction (Ga cohort) and internal validate (Gb cohort)of the model. Another cohort (I cohort) with 344 patients from Quzhou People's Hospital and 430 patients from Jiaxing Second People's Hospital was used for external validation. Univariate and multivariate binary logistic regression analyses were performed to identify the independent predictors. Then, the selected predictors were then used to establish the predictive nomogram. The apparent performance of the model was evaluated via externally validated. Decision curve analysis was also performed to assess the clinical utility of the developed model. Univariate and multivariate logistic regression analyses showed that alkaline phosphatase (ALP), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), prostate specific antigen (PSA) and lactate dehydrogenase were independent predictors of IDC-P. Therefore, a predictive nomogram of IDC-P was constructed. The nomogram had a good discriminatory power (AUC = 0.794). Internal validation (AUC = 0.819)and external validation (AUC = 0.903) also revealed a good predictive ability. Calibration curves showed good agreement between the predicted and observed incidences of IDC-P. We developed a clinical predictive model composed of alkaline phosphatase (ALP), total cholesterol (TC), triglycerides (TG), high-density lipoprotein (HDL), prostate specific antigen (PSA) and lactate dehydrogenase (LDH) with a high precision and universality. This model provides a novel calculator for predicting the diagnosis of IDC-P and different treatment options for patients at an early stage.

S41 Serum Lactate Dehydrogenase Levels: The Grim Reaper Sign in Acute Pancreatitis?

Introduction: Acute pancreatitis (AP) is one of the most common gastrointestinal presentations to the emergency department. Current scoring systems in place for predicting the severity and mortality of AP include Ranson criteria, the Acute Physiology And Chronic Health Evaluation II (APACHE II), and the Bedside Index of Severity in Acute Pancreatitis (BISAP) score. However, these scoring systems have their limitations and variable utility. Lactate dehydrogenase (LDH) is an enzyme found in almost all body tissues that is released during tissue damage and can be used as a marker of an organ injury. The purpose of this study was to investigate whether specific LDH values can predict mortality risk in patients with AP and it’s correlation with hospital length of stay. Methods: We conducted a retrospective cohort study of patients who had presented to HCA Healthcare facilities with the diagnosis of AP in the period of January 2011 - January 2021. A Total 514 patients that presented with AP and had a serum LDH data obtained on admission were identified and divided into three groups based on LDH cutoff levels. Length of hospital stay, length of intensive care unit stay (ICU), and mortality rate were compared among groups. Other variables included age, gender, race, BMI, and prior medical history of heart disease or cerebrovascular accident. The study was conducted using a Logistic Regression approach that used statistical analysis to predict the odds of a desired association. Results: Total 514 eligible patients were identified and divided into three groups. Group 1 consisted of 301 patients with LDH level < 300 IU/L, Group 2 consisted of 158 patients with LDH level between 300-600 IU/L, and Group 3 consisted of 55 patients with LDH level >600. AP patients with an initial LDH value of more than 600 IU/L on admission were likely to have longer hospital length of stay, by 4.5 days on average, 3.2 times more likely to be admitted to ICU, and 12.1 time more likely to expire than patients with an initial LDH value of less than 300 IU/L. Conclusion: This study has demonstrated that serum LDH is a cheap and a convenient test that can be obtained in AP patients to predict the length of hospital stay, ICU needs, and mortality rates. However, given our study is retrospective, additional randomized controlled studies are necessary to corroborate the beneficial effects of obtaining LDH in patients presenting with AP on admission (Table). Table 1. - The average length of hospital stay, percentage of patients required ICU care, and percentage of patients who has expired in each group along with the median and mode Groups Average Length of Hospital Stay Percentage of Patients required ICU care Percentage of Patients who has expired Group 1 8.09 22.92% 3.32% Group 2 10.03 33.54% 10.76% Group 3 15.63 54.55% 23.64%

Discovery of novel human lactate dehydrogenase inhibitors: Structure-based virtual screening studies and biological assessment

Most cancer cells switch their metabolism from mitochondrial oxidative phosphorylation to aerobic glycolysis to generate ATP and precursors for the biosynthesis of key macromolecules. The aerobic conversion of pyruvate to lactate, coupled to oxidation of the nicotinamide cofactor, is a primary hallmark of cancer and is catalyzed by lactate dehydrogenase (LDH), a central effector of this pathological reprogrammed metabolism. Hence, inhibition of LDH is a potential new promising therapeutic approach for cancer. In the search for new LDH inhibitors, we carried out a structure-based virtual screening campaign. Here, we report the identification of a novel specific LDH inhibitor, the pyridazine derivative 18 (RS6212), that exhibits potent anticancer activity within the micromolar range in multiple cancer cell lines and synergizes with complex I inhibition in the suppression of tumor growth. Altogether, our data support the conclusion that compound 18 deserves to be further investigated as a starting point for the development of LDH inhibitors and for novel anticancer strategies based on the targeting of key metabolic steps.

Inhibition of LDHA to induce eEF2 release enhances thrombocytopoiesis

AbstractTranslation is essential for megakaryocyte (MK) maturation and platelet production. However, how the translational pathways are regulated in this process remains unknown. In this study, we found that MK/platelet–specific lactate dehydrogenase A (LdhA) knockout mice exhibited an increased number of platelets with remarkably accelerated MK maturation and proplatelet formation. Interestingly, the role of LDHA in MK maturation and platelet formation did not depend on lactate content, which was the major product of LDHA. Mechanism studies revealed that LDHA interacted with eukaryotic elongation factor 2 (eEF2) in the cytoplasm, controlling the participation of eEF2 in translation at the ribosome. Furthermore, the interaction of LDHA and eEF2 was dependent on nicotinamide adenine dinucleotide (NADH), a coenzyme of LDHA. NADH-competitive inhibitors of LDHA could release eEF2 from the LDHA pool, upregulate translation, and enhance MK maturation in vitro. Among LDHA inhibitors, stiripentol significantly promoted the production of platelets in vivo under a physiological state and in the immune thrombocytopenia model. Moreover, stiripentol could promote platelet production from human cord blood mononuclear cell–derived MKs and also have a superposed effect with romiplostim. In short, this study shows a novel nonclassical function of LDHA in translation that may serve as a potential target for thrombocytopenia therapy.

Differential Diagnosis of Seizure and Syncope by the Means of Biochemical Markers in Emergency Department Patients

Background:Seizure and syncope have similar clinical symptoms but different etiologies. Hence, differential diagnosis is crucial prior to intervention. This study evaluates the diagnostic importance of neuron specific enolase (NSE), creatine phosphokinase (CPK), and serum lactate dehydrogenase (LDH) for admitting patients with seizure medical history to emergency department (ED) in order for differential diagnosis between syncope and seizure.Methods:Patients with a short-lasting loss of consciousness admitted to the ED were recruited. All patients with a short-lasting loss of consciousness were eligible and EEG was conducted several times and was taken over a long period. Patients were then divided into two groups of seizure and syncope. The biochemical markers levels of all the eligible patients were measured by a reputable laboratory.Results:In order to define specificity and sensitivity of different levels of biomarkers and the optimal cut-off points, ROC curves for each biomarker of syncope and seizure patients admitted to ED were performed. AUC for NSE, CPK, and LDH were 0.973 ± 0.023, 0.827 ± 0.047, and 0.836 ± 0.043 respectively in 95% confidence level. Cut-off points for NSE, CPK, and LDH were determined 25.12, 218.09, and 193.88 respectively.Conclusions:It was concluded that NSE, CPK and LDH levels were different significantly in seizure patients compared to syncope ones. The seizure group showed an increase in NSE, CPK and LDH level.

MiR-195-5p Ameliorates Cerebral Ischemia-Reperfusion Injury by Regulating the PTEN-AKT Signaling Pathway

BackgroundMiR-195-5p has been shown to play crucial roles in tumor inhibition, but its biological functions in cerebral ischemia-reperfusion (I/R) injury are unclear.MethodsTo mimic cerebral I/R injury, mice were induced by transient middle cerebral artery occlusion (MCAO). Human brain microvascular endothelial cells (HBMVECs) were treated with oxygen-glucose deprivation (OGD) to mimic I/R injury in vitro. The expression of miR-195-5p and PTEN was detected by qRT-PCR or Western blot. Cell viability was evaluated by CCK-8 assay. Cell apoptosis was detected by flow cytometer. Cell death was detected using specific lactate dehydrogenase (LDH) cytotoxicity kit. Infarct volume in mice brains was evaluated by TTC staining. Histopathological analysis was performed by HE staining and TUNEL staining. The interaction between miR-195-5p and PTEN was determined by TargetScan and luciferase reporter assay.ResultsMiR-195-5p was significantly downregulated and PTEN was upregulated during cerebral I/R injury both in vitro and in vivo. Overexpression of miR-195-5p efficiently enhanced cell viability, while reduced LDH release and apoptotic rate of OGD-treated HBMVECs in vitro. MiR-195-5p could negatively regulate the expression of PTEN by directly binding to its 3′-UTR. Overexpression of PTEN obviously attenuated the protective effect of miR-195-5p mimics on cell viability, LDH release and apoptosis in OGD-treated HBMVECs. Meanwhile, overexpression of miR-195-5p increased the expression levels of p-AKT in OGD-treated HBMVECs, while this effect was reversed by overexpression of PTEN. Moreover, overexpression of miR-195-5p efficiently ameliorated brain injury of mice after MCAO treatment in vivo.ConclusionOverexpression of miR-195-5p ameliorated cerebral I/R injury by regulating the PTEN-AKT signaling pathway, providing a potential therapeutic target for cerebral I/R injury.

Association of detectable levels of circulating tumor DNA (ctDNA) with disease burden in prostate cancer (PC).

5562 Background: PC is characterized by a relatively low prevalence of recurrent somatic point mutations. ctDNA is shed from PC and can be analyzed to profile somatic point mutations and copy number changes. We evaluated a computational approach to detect ctDNA (ie. ctDNA+) in PC based on allele frequencies of polymorphisms and mutations. We then sought to confirm the association of this biomarker with disease burden and clinical outcome. Methods: Customized, hybrid capture, high-depth next-generation sequencing was performed on pre-treatment (PT) plasma samples from a phase 2 line 3+ metastatic castration-resistant PC (mCRPC) study (NCT02854436, GALAHAD) and PT and end of treatment (EOT) samples from randomized Phase 3 study in non-metastatic (nm) CRPC (NCT01946204, SPARTAN) and from metastatic castration-sensitive PC (mCSPC) (NCT02489318, TITAN). Associations of ctDNA+ with bone lesions (number), visceral metastases (+/-), circulating tumor cells count (CTCc), and serum prostate specific antigen (PSA), alkaline phosphatase (AP) and lactate dehydrogenase (LD) were tested. Also, associations of ctDNA+ with overall survival (OS) and second progression free survival (PFS2) were evaluated in randomized studies using Cox regression. Results: ctDNA+ at PT was 7.5% in nmCRPC, 23.7% in mCSPC and 66% in heavily pre-treated mCRPC. ctDNA+ increased from PT to EOT in nmCRPC (7.5% to 27%) and mCSPC (23.7% to 63.6%). Disease burden metrics were evaluated in ctDNA+ vs ctDNA- patients. ctDNA+ was associated with higher disease burden in mCRPC (Table), nmCRPC and mCSPC. At EOT, ctDNA+ patients had shorter OS and PFS2 in nmCRPC (HR [95% CI] OS: 2.73 [1.83, 4.08], p &lt; 0.0001; PFS2: 2.00 [1.38, 2.90], p = 0.0002) and mCSPC (HR [95% CI] OS: 7.59 [3.22, 17.91], p &lt; 0.0001; PFS2: 4.84 [2.47, 9.47], p &lt; 0.0001). Conclusions: ctDNA+ assessed using our novel, composite biomarker increases with advanced disease state and disease progression, is significantly associated with disease burden and poor clinical outcome in PC and could be a clinically relevant metric for monitoring response to therapy. Clinical trial information: NCT02854436 . [Table: see text]

Differential diagnosis of tuberculous and malignant pleural effusions: comparison of the Th1/Th2 cytokine panel, tumor marker panel and chemistry panel

The differentiation between tuberculous plural effusion (TPE) and malignant plural effusion (MPE) remains a major clinical challenge in the diagnosis and management of pleural effusions, especially in developing countries with a high incidence of tuberculosis. We aimed to evaluate the diagnostic value of cytokines, tumor markers and biochemical markers in the differentiation of TPE and MPE. Two hundred and forty-two patients were included, of whom 134 were diagnosed with MPE and 108 were diagnosed with TPE. In total, 12 markers were tested in pleural effusion samples from all subjects: Interleukin-2 (IL-2), Tumor necrosis factor alpha (TNF-α), Interferon (IFN)-γ, interleukins-4, 6, 10 (IL-4,6,10), cytokeratin-19 fragment (CYFRA 21-1), carcinoembryonic antigen (CEA), neuron specific enolase (NSE), adenosine deaminase (ADA), lactate dehydrogenase (LDH) and high sensitivity C- reactive protein (Hs-CRP). The diagnostic value of each marker was evaluated and compared by receiver operating characteristic (ROC) curves. In the 12 markers evaluated, TNF-α, IFN-γ, IL-6, CYFRA 21-1, CEA, ADA and Hs-CRP were significantly different between the TPE and MPE groups, and the areas under the ROC curves were 0.624, 0.942, 0.619, 0.808, 0.903, 0.842 and 0.917, respectively. IFN-γ showed a better diagnostic performance than the other markers. With a cut-off value of >2.45 pg/mL, the sensitivity and specificity of IFN-γ were 91.11 and 91.94%, respectively. TNF-α, IFN-γ, IL-6, CYFRA 21-1, CEA, ADA and Hs-CRP were useful in the differentiation between the TPE and MPE groups. IFN-γ showed a better diagnostic performance than the multitude of other markers evaluated in this study, which is satisfactory for the discrimination of TPE and MPE.

Consequences of a Metabolic Glucose-Depletion on the Survival and the Metabolism of Cultured Rat Astrocytes.

Brain astrocytes are considered to be highly glycolytic, but these cells also produce ATP via mitochondrial oxidative phosphorylation. To investigate how a metabolic depletion of glucose will affect the metabolism of astrocytes, we applied glucose at an initial concentration of 2mM to cultured primary astrocytes and monitored the cell viability and various metabolic parameters during an incubation for up to 2weeks. Already within 2days of incubation the cells had completely consumed the applied glucose and lactate had accumulated in the medium to a concentration of around 3mM. During the subsequent 10days of incubation, the cell viability was not compromised while the extracellular lactate concentration declined to values of around 0.2mM, before the cell viability was compromised. Application of known inhibitors of mitochondrial metabolism strongly accelerated glucose consumption and initial lactate production, while the lactate consumption was completely (antimycin A or 8-hydroxy efavirenz) and partially (efavirenz, metformin or tyrphostin 23) inhibited which caused rapid and delayed cell toxicity, respectively. The switch from glycolytic glucose metabolism to mitochondrial metabolism during the incubation was neither accompanied by alterations in the specific cytosolic lactate dehydrogenase activity or in the WST1 reduction capacity nor in the mitochondrial citrate synthase activity, but a cellular redistribution of mitochondria from a perinuclear to a more spread cytoplasmic localization was observed during the lactate consumption phase. These results demonstrate that cultured astrocytes survive a metabolism-induced glucose depletion very well by consuming lactate as fuel for mitochondrial ATP generation.

Relationship between serum markers and volume of liver metastases in castration-resistant prostate cancer

BackgroundProstate cancer patients with liver metastases have a poor prognosis. To date, no study exists investigating the relationship between liver tumor burden and clinical laboratory markers. Materials and MethodsMetastatic castrate-resistant prostate cancer (mCRPC) patients with radiographic evidence of liver metastases were selected for this study. Volumetric measurements of liver metastases were ascertained for all available patients. Prostate specific antigen (PSA), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), albumin (ALB), total bilirubin and hemoglobin (HGB) levels were then assessed to coincide with the scan dates. Univariate and multivariate mixed-model regression analysis were performed to evaluate the relationship between laboratory markers and liver lesion volume. Data sets with non-normal distribution were logarithmically transformed. Akaike information criteria (AIC) was used to identify the most reliable multivariate model. ResultsIn our heavily pretreated liver-metastatic patient population, univariate analysis demonstrated a statistically significant positive correlation between PSA (p = 0.0002), ALP (p = 0.0305), AST (p < 0.0001), ALT (p = 0.0049), and LDH (p = 0.0019) and liver lesion volume. Additionally, ALB (p = 0.0006) and HGB (p = 0.0103) had statistically significant negative correlation. Multivariate analysis identified AST and hemoglobin assessments as the best predictors of increasing liver lesion burden. Preliminary data on circulating tumor DNA (ctDNA) mutational and amplification findings are also reported. ConclusionsAnalysis identified AST and hemoglobin as optimal predictors of liver lesion volume. These patients have a heavy burden of ctDNA abnormalities. Further studies with a larger patient population are needed to verify these results.Micro Abstract: This study investigates the association between liver lesion burden and clinical laboratory markers in castrate-resistant prostate cancer patients with hepatic metastases. Our univariate analysis identified multiple laboratory markers as significant indicators of worsening hepatic disease. Multivariate analysis demonstrated that AST and hemoglobin were the most effective predictors of change in liver lesion volume.

The use of Fionet technology for external quality control of malaria rapid diagnostic tests and monitoring health workers' performance in rural military health facilities in Tanzania.

IntroductionInternal and external quality control (QC) of rapid diagnostic tests (RDTs) is important to increase reliability of RDTs currently used to diagnose malaria. However, cross-checking of used RDTs as part of quality assurance can rarely be done by off-site personnel because there is no guarantee of retaining visible test lines after manufacturers’ recommended reading time. Therefore, this study examined the potential of using Fionet™ technology for remote RDT quality monitoring at seven clinics, identifying reasons for making RDT processing and interpretation errors, and taking corrective actions for improvement of diagnosis and consequently improved management of febrile patientsMethodsThe study was conducted at seven military health facilities in Mainland Tanzania and utilized RDTs capable of detecting Plasmodium falciparum specific Histidine-rich protein 2 (Pf-HRP2) and the genus specific Plasmodium lactate dehydrogenase (pLDH) for other species of plasmodium (P. vivax, P. malariae or P. ovale; pan-pLDH). Patients’ data and images of processed RDTs from seven clinics were uploaded on a Fionet web portal and reviewed regularly to monitor preparation procedures and visual interpretation of test results compared to automated analysis using the Deki reader of RDT. Problems detected were rapidly communicated to remote laboratory personnel at the clinic for corrective action and follow-up of patients who were falsely diagnosed as negative and missed treatment. Factors contributing to making errors in visual interpretation of RDT results were analyzed during visits to the health facilities.ResultsA total of 1,367 (1.6%) out of 83,294 RDT test images uploaded to the Fionet portal had discordant test results of which 822 (60.1%) and 545 (39.9%) were falsely reported as negative and positive, respectively. False negative and false positive test results were common for a single test line in 515 (62.7%) and 741 (54.2%) tests, respectively. Out of 1,367 RDT images assessed, 98 (7.2%) had quality problems related to preparation procedures of which 95(96.9%) errors were due to putting too much blood on the sample well or insufficient buffer in the respective wells. The reasons for discrepant results included, false reporting of none existent lines in 526 (38.5%) tests, missing a faint positive line in 493 (36.1%), missing a strong positive line in 248(18.1%) and errors caused by poorly processed RDTs in 96 (7.2%) tests. Among the false negative tests (n = 822), 669 (48.9%) patients were eligible for follow–up and only 339 (48.5%) were reached and 291 (85.8%) received appropriate anti-malaria therapy.ConclusionFionet technology enabled remote monitoring of RDT quality issues, identifying reasons contributing to laboratory personnel making errors and provided a rapid method to implement corrective actions at remote sites to improve malaria diagnosis and consequently improved health care management of febrile patients infected with malaria.

Inhibition of lactate dehydrogenase A suppresses inflammatory response in RAW 264.7 macrophages.

Lactate is an important metabolite in cellular metabolism and fluctuates in certain disease conditions including cancer and immune diseases. It was hypothesized that a decrease in lactate would modulate the inflammatory response elicited by lipopolysaccharides (LPS) in macrophages. When RAW 264.7 macrophages were treated with FX11, a specific lactate dehydrogenase (LDHA) inhibitor, the expression of the cytokines, inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX‑2) was downregulated due to reduced cellular lactate levels. Genetic suppression of LDHA by small interfering RNA (siRNA) downregulated the LPS‑activated expression of interleukin (IL)‑6, iNOS, and COX‑2, and reduced the production of IL‑6 and nitrites. Pharmacological and genetic suppression of LDHA inhibited the phosphorylation of p38 mitogen‑activated protein kinase. Microarray gene expression profile demonstrated that the genes involved in cell proliferation and inflammation were mainly altered by siRNA‑mediated LDHA suppression. Collectively, the present observations suggest that lactate may be an important metabolite and implicated in regulation of inflammatory response.

Functional and Structural Resilience of the Active Site Loop in the Evolution of Plasmodium Lactate Dehydrogenase.

The malarial pathogen Plasmodium falciparum ( Pf) is a member of the Apicomplexa, which independently evolved a highly specific lactate dehydrogenase (LDH) from an ancestral malate dehydrogenase (MDH) via a five-residue insertion in a key active site loop. PfLDH is widely considered an attractive drug target because of its unique active site. The conservation of the apicomplexan loop suggests that a precise insertion sequence was required for the evolution of LDH specificity. Aside from a single critical tryptophan, W107f, the functional and structural roles of residues in the loop are currently unknown. Here we show that the loop is remarkably robust to mutation, as activity is resilient to radical perturbations of both loop identity and length. Thus, alternative insertions could have evolved LDH specificity as long as they contained a tryptophan in the proper location. PfLDH likely has great potential to develop resistance to drugs designed to target its distinctive active site loop.

Predictor of 68Ga PSMA PET/CT positivity in patients with prostate cancer.

The aim of this study was to evaluate predictive factors of 68Gallium (68Ga) prostate-specific membrane antigen (PSMA) positron emission tomography (PET)/computed tomography (CT) positivity. Relationships between serum prostate specific antigen (PSA), lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) levels, Gleason Score (GS) and positivity of 68Ga PSMA PET in patients who underwent 68Ga PSMA PET/CT for restaging for PCa were evaluated retrospectively. One hundred and four (median age: 67; range: 51-88) patients were included in this study. Of these patients, PSMA PET was positive in 75 (72%) patients. Mean serum PSA levels for PET negative and positive groups were 0.76±1.00 and 180.85±324.93 ng/mL (P<0.001). The sensitivity and specificity of 68Ga PSMA PET/CT for detection of disease recurrence were calculated as 92% and 80%, respectively, for the 1.4 ng/mL PSA cut-off and 92% and 90%, respectively, for the 2 ng/mL PSA cut-off values. The positivity rates for patients with PSA levels <1.4 ng/mL and ≥1.4 ng/mL were 21% and 90%, respectively (P<0.001). 68Ga PSMA PET/CT seems to be a highly sensitive in patients with early PSA recurrence. Patients with higher GS and early PSA recurrence could benefit from 68Ga PSMA PET/CT.

Antimalarial activity of phytol derivatives: in vitro and in vivo study

Severe malaria is one of the leading causes of mortality among children and pregnant woman globally. Resistance development against the frontline antimalarial drugs has created an alarming situation, which requires intensive drug discovery to develop effective, affordable, and accessible antimalarial agents possessing novel modes of action. As a part of our drug discovery program for antimalarial agents from plants, phytol a very commonly occurring diterpene alcohol in the plant was investigated for its antimalarial potential. In vitro antiplasmodial activity against the chloroquine-sensitive Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase (pfLDH), showed moderate activity (IC50 211.5 ± 0.93 µM). Further, phytol was chemically converted into thirteen derivatives, which were evaluated for their antiplasmodial potential. All the derivatives were moderate active, but among all the derivatives palmitoyl (PhY-3; IC50 12.13 ± 0.31 µM) has found most active without any cytotoxic effect on macrophage cells. PhY-3 was further validated in an in vivo system for its efficacy and safety profile in mice. Oral administration of PhY-3 showed significant reduction of parasitemia and increased mean survival time. It also attributed significant increase in blood glucose and hemoglobin level, when compared with vehicle-treated P. berghei infected mice without any toxic effect on normal mice at a higher dose. These findings confirm suitability of the phytol derivatives as new chemical entities for further investigation towards the management of malaria.

Salivary Enzymological Profile Of Lactate Dehydrogenase (LDH), Tartrate Resistance Acid Phosphatase (TRAP), Alkaline Phosphatase (ALP) And Aspartate Aminotransferase (AST) During Orthodontic Treatment

Introduction: Orthodontic tooth movement is a complex process involving tooth and periodontal tissue, which release enzymes and biomarkers. The aim of this study was to investigate enzymes activities of salivary fluid during orthodontic treatment. Materials and Methods: A group of nineteen healthy subject (mean age 21.5 years) with Class II/1 malocclusion who required extraction of maxillary first premolar were recruited. Saliva samples were collected from these 19 patients that underwent canine traction using fixed appliances. Enzymes activities were measured before placement of fixed appliances (basal activity) followed with immediately before and weekly canine retraction for five weeks. The specific Lactate Dehydrogenase (LDH), Aspartate Aminotransferase (AST), Tartrate Resistance Acid Phosphatase (TRAP) and Alkaline Phosphatase (ALP) activities in saliva sample were analyzed using spectrophotometer (405nm). Result: No statistical significant (p&gt; 0.05) difference was noted in LDH activity between basal activity and during canine retraction period. AST showed higher activity compared to basal activity from week 0 to week 1 with statistically significant increased (p&lt;0.05) found in week 1. ALP showed significantly higher enzyme activity compared to basal activity from week 1 to 5, with the peak level at week 5. While TRAP showed significant increase in enzyme activity compared to basal activity only at week 2 after canine retraction. Conclusion(s): Orthodontic tooth movement can be monitored through the expression of AST, TRAP and ALP activity in saliva. Saliva LDH cannot be used as a biomarker in monitoring tooth movement.