Specific Lactate Dehydrogenase Research Articles

The prognostic value of the serum neuron specific enolase and lactate dehydrogenase in small cell lung cancer patients receiving first-line platinum-based chemotherapy.

The aim of this study was to investigate the associations of serum levels of neuron-specific enolase (NSE), pro-gastrin releasing peptide (ProGRP), and lactate dehydrogenase (LDH) with clinical response and survival in small cell lung cancer (SCLC) patients receiving first-line platinum-based chemotherapy.One hundred thirty-six patients with SCLC were recruited in this study. All the patients received first-line platinum-based chemotherapy. Clinical efficacy was assessed according to Response Evaluation Criteria in Solid Tumors v1.1 criteria. Serum samples were collected from SCLC patients before chemotherapy. NSE, ProGRP, and LDH levels were measured by commercial electrochemiluminescence immunoassay, enzyme-linked immune sorbent assay, and kinetic spectrophotometric method, respectively.Overall response rate was 71.3% with 97 patients who achieved complete response (CR) + partial response (PR). NSE and LDH level declined in patients who achieved CR + PR compared with patients in stable disease (SD) and progress disease (PD). Multivariate logistic regression analysis revealed that NSE > 50.324 ng/mL, stage ED, and distant metastases were independent risk factors for patients achieving CR + PR, and chemotherapy > 4 cycles was an independent protective factor in predicting CR + PR. Receiver operating characteristic (ROC) curves presented that expression of NSE, ProGRP, and LDH are of good predicting value for patients achieving CR + PR. Patients with a higher level of NSE and LDH presented worse progression-free survival and overall survival. In addition, multivariate Cox regression analysis showed that NSE level > 50.324 ng/mL and distant metastasis were independently correlated with worse OS.Serum NSE and LDH could be promising biomarkers for predicting therapy response and survival of SCLC patients receiving first-line platinum-based chemotherapy.

Effects of Lactate Dehydrogenase (LDH) in Preeclampsia

Background: Lactate dehydrogenase (LDH) is a multifaceted enzyme whose effects in pregnancy related complications e.g., preeclampsia (PE) is now gaining attention. Here we present evidence that LDH levels could contribute significantly to the outcomes of PE. Aim: To determine the effects of LDH in PE. Methodology: The case control study was located at the Obstetrics and Gynaecology department of the KorleBu Teaching Hospital (KBTH). STROBE consensus checklist was adopted. One hundred and forty (140) consented subjects were recruited after ethical clearance was obtained and structured questionnaire administered to them. Four (4) mL blood and 5 mL urine samples were taken for biochemical analysis and urinalysis respectively. Randox and Sysmex automated chemistry analyser was used to quantify blood chemistry. The data was captured as protected health information (PHI) and analysed with SPSS version 22. Results: LDH exposure was associated with higher odds of outcome in preeclampsia (PE) [OR(CI)=4.76(1.26-18.72); p-value=0.0068]. However, with an adjusted OR, LDH categories were associated with birth weight. Notwithstanding the added input, in preeclampsia, increased LDH at <34 weeks of gestation related with decreased birthweight only when platelet, diastolic blood pressure (DBP), pH, bilirubin, parity and liver enzymes each served as covariates in the log linear logit analysis. Conclusion: LDH was associated with low birth weight in PE in a concentration dependent manner under the influence of predictors like pH, platelet and diastolic pressure (DBP) for causality. Therefore, a thoughtful planned foetal delivery under a specific LDH threshold and a regular monitoring of urine pH, full blood count (FBC), and blood pressure might improve the outcomes of PE

Diarylheptanoids Rich Fraction ofAlnus nepalensisAttenuates Malaria Pathogenesis:In-vitroandIn-vivoStudy

Diarylheptanoids from Alnus nepalensis leaves have been reported for promising activity against filariasis, a mosquito-borne disease, and this has prompted us to investigate its anti-malarial and safety profile using in-vitro and in-vivo bioassays. A. nepalensis leaf extracts were tested in-vitro against chloroquine-sensitive Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. Among all, the chloroform extract (ANC) has shown promising anti-plasmodial activity (IC50 8.06 ± 0.26 µg/mL). HPLC analysis of ANC showed the presence of diarylheptanoids. Efficacy and safety of ANC were further validated in in-vivo system using Plasmodium berghei-induced malaria model and acute oral toxicity in mice. Malaria was induced by intra-peritoneal injection of P. berghei infected red blood cells to the female Balb/c mice. ANC was administered orally at doses of 100 and 300 mg/kg/day following Peter's 4 day suppression test. Oral administration of ANC showed significant reduction of parasitaemia and increase in mean survival time. It also attributed to inhibition of the parasite induced pro-inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle-treated infected mice. In-vivo safety evaluation study revealed that ANC is non-toxic at higher concentration. Copyright © 2016 John Wiley & Sons, Ltd.

Acute kidney injury in patients with Plasmodium vivax malaria: Clinicohistopathological profile

Context: There has been an unexplained increase in the number of cases with multiorgan dysfunction including acute kidney injury (AKI), attributed to Plasmodium vivax monoinfection. Only a few case reports in literature have published the renal biopsy findings in these patients. Aims: The aim of this study was to evaluate the clinical and histopathologic profile of patients with P. vivax malaria monoinfection and AKI. Settings and Design: A prospective study was performed in a tertiary care hospital in North-Western India. Subjects and Methods: The study included patients diagnosed with P. vivax monoinfection on peripheral smear blood films and rapid diagnostic test (positive for P. vivax specific lactate dehydrogenase). AKI was defined based on the WHO criteria for complicated malaria, i.e. serum creatinine >265 μmol/l or 3 mg/dl. The patients were initiated on hemodialysis for persistent hyperkalemia, fluid overload, severe metabolic acidosis, or uremic symptoms. Renal biopsy was performed in the presence of active urinary sediments (proteinuria, hematuria) or persistence of renal failure >14 days. Results: A total of thirty patients fulfilled AKI criteria. The patients with AKI were older (mean age 42.1 ± 10.9 years), male, with a longer duration of illness (mean 12.3 ± 10 days) and associated with multisystem dysfunction. The mean serum creatinine was 7.58 ± 3.2 mg/dl, thrombocytopenia was seen in 47%. Thirty percent had severe anemia requiring a blood transfusion. Renal biopsy was performed in six patients for various indications. The most common pattern was acute tubular necrosis (four patients), followed by acute cortical necrosis (1), and thrombotic microangiopathy (one patient). The complete renal recovery was seen in 24 (80%). Two patients became dialysis-dependent. Conclusions: AKI associated with P. vivax monoinfection is not rare as previously thought. Therefore, it should be considered in the differential diagnosis of any patient presenting with AKI.

Present, Emerging and Possible Future Biomarkers in Castration Resistant Prostate Cancer (CRPC).

Recently, an increasing number of systemic therapies with life extending capacity have become available in metastatic castration resistant prostate cancer (mCRPC) i.e. Abiraterone acetate, Enzalutamide, Sipuleucel-T, Docetaxel, Cabazitaxel and Radium-223. More compounds are currently being evaluated in promising pivotal trials (e.g. Tasquinimod, ARN-509, ODM-201, and more). Limitations of the currently available biomarkers make treatment decisions challenging. Considering the ever increasing complexity of treatment algorithms in mCRPC the current demand of research is to find and characterize biomarkers with prognostic, predictive and surrogate quality, allowing for information on clinically meaningful outcomes and on which therapy to offer patients in different and complex scenarios. A comprehensive English-language literature review was performed through PubMed to identify articles and abstract presentations of the major conferences on cancer during December 2014. In this review we address established biomarkers like prostate specific antigen, lactate dehydrogenase and alkaline phosphatase. Emerging biomarkers like circulating tumor cells, androgen receptor splice variants, cancer stem cells and imaging biomarkers have also been reviewed and placed in the context of prognostic, predictive and surrogate implications in the current field of CRPC. We elaborate on the requirements of good biomarkers and discuss possible future developments of biomarkers in CRPC. Advances in knowledge of biomarkers in CRPC and thus biomarker driven therapy monitoring and up-front therapy decisions may help in the future to tailor treatment algorithms, give information on which therapy to offer and when to continue a given therapy in equivocal scenarios. This could maximize treatment benefit and minimize toxicity.

Lab on a chip for multiplexed immunoassays to detect bladder cancer using multifunctional dielectrophoretic manipulations.

A multiplexed immunosensor has been developed for the detection of specific biomarkers Galectin-1 (Gal-1) and Lactate Dehydrogenase B (LDH-B) present in different grades of bladder cancer cell lysates. In order to immobilize nanoprobes with different antibodies on a single chip we employed three-step programmable dielectrophoretic manipulations for focusing, guiding and trapping to enhance the fluorescent response and reduce the interference between the two antibody arrays. The chip consisted of a patterned indium tin oxide (ITO) electrode for sensing and a middle fish bone shaped gold electrode for focusing and guiding. Using ITO electrodes for the sensing area can effectively eliminate the background noise of fluorescence response as compared to metal electrodes. It was also observed that the three step manipulation increased fluorescence response after immunosensing by about 4.6 times as compared to utilizing DEP for just trapping the nanoprobes. Two different-grade bladder cancer cell lysates (grade I: RT4 and grade III: T24) were individually analyzed for detecting the protein expression levels of Gal-1 and LDH-B. The fluorescence intensity observed for Gal-1 is higher than that of LDH-B in the T24 cell lysate; however the response observed in RT4 is higher for LDH-B as compared to Gal-1. Thus we can effectively identify the different grades of bladder cancer cells. In addition, the platform for DEP manipulation developed in this study can enable real time detection of multiple analytes on a single chip and provide more practical benefits for clinical diagnosis.

Immunological Testing Reveals Exposure to Malaria in the Hypoendemic Region of Iran.

Background. South eastern parts of Iran remain endemic for malaria infection. There is some concern that malaria infection may spread into Bushehr, which is located in the south western part bordering the Persian Gulf and at the periphery of the declared endemic region Hormozgan province due to frequency of visitors from eastern endemic areas and from neighboring malaria endemic countries. We investigated malaria prevalence in Bushehr. Methods and Results. Attempts were made to identify malaria active infection in blood smears and malaria specific antibody and antigens in serum samples. Traditional blood smears prepared from 1955 blood specimens yielded no definitive malaria positive case by microscopic technique. A total of 270 (13.8%) serum samples were positive for malaria antibodies. Using specific ELISA kits, presence of histidine rich proteins and lactate dehydrogenase antigens were investigated in serum samples. No histidine rich proteins specific for P. falciparum were detected amongst 270 antibody positive samples. However, six samples representing 0.3% of total population, were found to be positive for plasmodium pan specific lactate dehydrogenase antigens. This suggested the possibility of low level exposure to malaria in Bushehr community. Conclusions. Out of a total of 1955 samples tested, 270 (13.8%) were positive for malaria antibodies and six (0.3%) of these were positive for plasmodium-specific lactate dehydrogenase antigen suggesting a low level exposure to malaria in a hypoendemic region based on immunological testing. Since none of the 270 antibody samples were positive for histidine rich protein antigens, there is scope for further testing of blood samples by molecular methods such as polymerase chain reactions to confirm the plasmodium species and provide information valuable for future investigations. Our testing strategy for hypoemdemic malaria can be used as a template for investing malaria in 32 eliminating countries for testing ongoing transmission. This approach may be useful as a method in epidemiological studies.

Plasmodium falciparum and Plasmodium vivax specific lactate dehydrogenase: Genetic polymorphism study from Indian isolates

Control and eradication of malaria is hindered by the acquisition of drug resistance by Plasmodium species. This has necessitated a persistent search for novel drugs and more efficient targets. Plasmodium species specific lactate dehydrogenase is one of the potential therapeutic and diagnostic targets, because of its indispensable role in endoerythrocytic stage of the parasite. A target molecule that is highly conserved in the parasite population can be more effectively used in diagnostics and therapeutics, hence, in the present study polymorphism in PfLDH (Plasmodiumfalciparum specific LDH) and PvLDH (Plasmodiumvivax specific LDH) genes was analyzed using PCR-single strand confirmation polymorphism (PCR-SSCP) and sequencing. Forty-six P. falciparum and thirty-five P. vivax samples were screened from different states of India. Our findings have revealed presence of a single PfLDH genotype and six PvLDH genotypes among the studied samples. Interestingly, along with synonymous substitutions, nonsynonymous substitutions were reported to be present for the first time in the PvLDH genotypes. Further, through amino acid sequence alignment and homology modeling studies we observed that the catalytic residues were conserved in all PvLDH genotypes and the nonsynonymous substitutions have not altered the enzyme structure significantly. Evolutionary genetics studies have confirmed that PfLDH and PvLDH loci are under strong purifying selection. Phylogenetic analysis of the pLDH gene sequences revealed that P. falciparum compared to P. vivax, has recent origin. The study therefore supports PfLDH and PvLDH as suitable therapeutic and diagnostic targets as well as phylogenetic markers to understand the genealogy of malaria species.

Antimalarial activity of Evolvulus alsinoids Linn.-an in vitro Plasmodium falciparum specific lactate dehydrogenase enzyme inhibition assay

ObjectiveTo investigate the effect of standardized methanol extract of Evolvulus alsinoids Linn. (E. alsinoids) on Plasmodium falciparum specific lactate dehydrogenase (PfLDH) enzyme inhibition. MethodsTo carry out enzyme inhibition studies, lactate dehydrogenase was cloned from Plasmodium falciparum 3D7 strain using expression vector pET28a and expressed in Escherichia coli BL21 (DE3). Protein purification was carried out by Ni-affinity chromatography. This protein was used for the inhibition study. Methanol extract of E. alsinoids was standardized by high performance thin layer chromatography described by previous literature and screened for PfLDH enzyme inhibitory activity and compared with gossypol (a known PfLDH enzyme inhibitor). ResultsIt was found that E. alsinoids possesses a compond scopoletin and potentially inhibits PfLDH [(25.04依0.51)%]. ConclusionsMethanol extract of E. alsinoids showed significant PfLDH inhibition as evidenced from the experiments performed. The activity may be attributed to the presence of various polyphenolic and flavanoids compounds.

The Antinociceptive Effect of Dexmedetomidine Modulates Spleen Cell Immunity in Mice

Background: Pain plays roles in both the nervous system and immune system. Changes in the neuroendocrine pathway under pain conditions give rise to sympathetic outflow with increased plasma catecholamines and activate immune reactions. Dexmedetomidine exerts sedative, analgesic, and anesthetic-sparing effects and is known to diminish pro-inflammatory processes by central sympatholytic effects. To investigate the influence of the analgesic effect of dexmedetomidine on immunomodulation under pain conditions, splenic natural killer (NK) tumoricidal cytotoxic activity, proliferative ability of T lymphocytes, and cytokine changes were assessed.Methods: After evaluation of the analgesic efficacy of dexmedetomidine in C57BL mice that were subjected to formalin-induced pain, dexmedetomidine (30 µg/kg) or saline was injected intraperitoneally (ip) 30 min before formalin (20 µL of 2% formalin in 0.9% saline) injection. NK cell activity against NK-sensitive YAC-1 lymphoma cells was evaluated by the percentage of specific lactate dehydrogenase (LDH) release. Various numbers of effector cells (NK cells) were added to the wells of a microtiter plate containing 2 × 104 target YAC-1 cells in 100 μL, to achieve final effector-to-target cell ratios of 80:1, 40:1, and 20:1. The level of lymphocyte proliferation in response to phytohemagglutinin (PHA) was detected by bromodeoxyuridine (BrdU) incorporation assay. TNF-α, IL-1β, and IL-10 levels were determined in blood samples and supernatants of splenocyte preparations.Results: IP administration of dexmedetomidine significantly decreased the time of licking and biting during the first and second phases of the formalin test (p <0.001). Formalin-induced pain led to higher activity of NK cells than in sham-treated mice (p <0.05), but NK activity was not increased significantly by ip dexmedetomidine treatment. Formalin-induced pain significantly increased splenic lymphocyte proliferation (p <0.05), but dexmedetomidine did not alter this response. There was a significant increase in plasma TNF-α (p = 0.048) and IL-6 (p = 0.014) levels after formalin-induced pain. However, the differences between the responses after ip dexmedetomidine did not change significantly.Conclusions: Dexmedetomidine showed antinociceptive effect on both of acute pain phase 1 and hyperalgesic phase 2 of formalin pain model. Formalin-induced pain alters cellular immunity of spleen in mice. Dexmedetomidine attenuates the activation of NK cells under pain condition, but neither the proliferative response of the splenic lymphocytes nor the cytokine production was affected by dexmedetomidine.

Stable Suppression of Lactate Dehydrogenase Activity during Anoxia in the Foot Muscle ofLittorina littoreaand the Potential Role of Acetylation as a Novel Posttranslational Regulatory Mechanism

The intertidal marine snail, Littorina littorea, has evolved to withstand extended bouts of oxygen deprivation brought about by changing tides or other potentially harmful environmental conditions. Survival is dependent on a strong suppression of its metabolic rate and a drastic reorganization of its cellular biochemistry in order to maintain energy balance under fixed fuel reserves. Lactate dehydrogenase (LDH) is a crucial enzyme of anaerobic metabolism as it is typically responsible for the regeneration of NAD+, which allows for the continued functioning of glycolysis in the absence of oxygen. This study compared the kinetic and structural characteristics of the D-lactate specific LDH (E.C. 1.1.1.28) from foot muscle of aerobic control versus 24 h anoxia-exposed L. littorea. Anoxic LDH displayed a near 50% decrease in V max (pyruvate-reducing direction) as compared to control LDH. These kinetic differences suggest that there may be a stable modification and regulation of LDH during anoxia, and indeed, subsequent dot-blot analyses identified anoxic LDH as being significantly less acetylated than the corresponding control enzyme. Therefore, acetylation may be the regulatory mechanism that is responsible for the suppression of LDH activity during anoxia, which could allow for the production of alternative glycolytic end products that in turn would increase the ATP yield under fixed fuel reserves.

Correlations among homocysteine, cardiac troponin I and cardiac enzymes in different ages of clinically healthy male dromedary camels

BACKGROUND: Information regarding serum biochemical profile can reflect cardiovascular performance in animals. Although studies have evaluated the inter-relationship among cardiovascular biomarkers in animals and human beings, there are no reports of such a probable relationship in camelids. OBJECTIVES: The aim of the present study was to provide data on the correlations among cardiovascular biomarkers in different ages of clinically healthy male dromedary camels to provide a basis for assessing cardiac muscle healthiness in this species. METHODS: Thirty clinically healthy dromedary camels (Camelus dromedarius) were selected and divided into four age groups including 1-3 (n=7), 4-6 (n=7), 7-9 (n=8), and above 10 (n=8) years old. Blood samples were collected and sera were separated. Serum concentrations of homocysteine (Hcy), cardiac troponin I (cTnI), creatine kinase-myocardial specific isoenzymes (CK-MB), lactate dehydrogenase (LDH), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were evaluated. RESULTS: The results of the present study showed that there were significant correlations among cTnI and CK-MB (r=-0.853; p=0.015) and Hcy (r=0.916; p=0.004) in the 4 to 6-years-old group of clinically healthy male dromedary camels. LDH was significantly correlated with CK-MB in the 7 to 9-year-old group (r=-0.710; p=0.045). There were no significant correlations among different factors of 1-3 and above 10-year-old groups (p>0.05). CONCLUSIONS:The data provided here is the first report on cardiac health assessment parameters in dromedary camels. Moreover, the data is valuable in camel racing clubs, when an overall cardiac health and fitness is to be assessed. The correlation reported here might also be helpful for easier analysis of cardiac health status in dromedary camels. The data may be useful for assessing suspected cases of myocardial diseases and its changes maybe of prognostic value.

Antimalarial and safety evaluation of Pluchea lanceolata (DC.) Oliv. & Hiern: In-vitro and in-vivo study

Ethnopharmacological relevanceMany of the effective therapeutic strategies have been derived from ethnopharmacologically used natural products. Pluchea lanceolata is an herb employed in Indian folk medicine for malaria like fever but it lacks proper pharmacological intervention. Aim of the studyTo evaluate antimalarial and safety profile of Pluchea lanceolata: an in-vitro, in-vivo for its ethnopharmacological validation. Materials and methodsMethanol, butanol, ethyl acetate, chloroform, hexane extracts and its isolate, taraxasterol acetate (TxAc) were obtained from air dried aerial part of Pluchea lanceolata. These were tested in-vitro against chloroquine-sensitive strain of Plasmodium falciparum NF54 by measuring the parasite specific lactate dehydrogenase activity. The most potent hexane extract and TxAc were further validated for in-vivo antimalarial and safety evaluation. TxAc, a pentacyclic-triterpene isolated from the most active fraction was further evaluated with special emphasis on inflammatory mediators involved in malaria pathogenesis. Murine malaria was induced by intra-peritoneal injection of Plasmodium berghei infected red blood cells to the male Swiss inbred mice. Mice were orally treated following Peters 4-Day suppression test. In-vivo antimalarial efficacy was examined by evaluating the parasitaemia, percent survival, mean survival time, blood glucose, haemoglobin and pro-inflammatory mediators involved in malaria pathogenesis. ResultsHexane extract and TxAc showed promising antimalarial activity in-vitro and in-vivo condition. TxAc attributed in inhibition of the pro-inflammatory cytokines as well as afford to significant increase in the blood glucose and haemoglobin level when compared with vehicle treated infected mice. We have not observed the synergistic action of combinations of chloroquine and TxAc from our experimental results. In-vitro and in-vivo safety evaluation study revealed that hexane extract is non toxic at higher concentration. ConclusionPresent study further validates the ancient Indian traditional knowledge and use of Pluchea lanceolata as an antimalarial agent. Study confirms the suitability of Pluchea lanceolata as a candidate for further studies to obtain a prototype for antimalarial medicine.

Effects of l-carnitine supplementation to suckling piglets on carcass and meat quality at market age

In a previous study, carnitine supplementation to piglets during the suckling period resulted in an increased total muscle fibre number at weaning in piglets of low birth weight. The objective of the present study was to investigate whether this effect is maintained until market age and whether this would attenuate the negative consequences of low birth weight on carcass and meat quality. Using a split-plot design with litter as block, sex as whole plot and treatment as subplot, the effects of early-postnatal l-carnitine supplementation on female and castrated male piglets of low birth weight were investigated on a total of 56 German Landrace piglets from 14 litters. From days 7 to 27 of age piglets were orally supplemented once daily with 400 mg of l-carnitine dissolved in 1 ml of water or received an equal volume of water without carnitine. From weaning (day 28) until slaughter (day 166 of age) all pigs were fed standard diets. At weaning, carnitine-supplemented piglets had a twofold increased concentration of free carnitine (P < 0.001) and a lower concentration of non-esterified fatty acids (P < 0.05) in blood plasma indicating that carnitine became bioavailable and increased fatty acid utilization during the period of supplementation. Growth performance was not influenced by treatment in any growth period. Dual-energy X-ray absorptiometry revealed no differences in body composition between groups in weeks 12, 16 and 20 of age. LW at slaughter, carcass weight, measures of meat yield and fat accretion, as well as body composition by chemical analyses and dissection of primal cuts did not differ between treatments. No differences between control and carnitine-treated pigs in total fibre number (P = 0.85) and fibre cross-sectional area (P = 0.68) in m. semitendinosus (ST) measured at slaughter could be observed. The carnitine group tended to exhibit a smaller proportion of slow-twitch oxidative fibres (P = 0.08), a greater proportion of fast-twitch glycolytic fibres (P = 0.11), and increased specific lactate dehydrogenase activity (P = 0.09) in ST indicating a more glycolytic muscle metabolism. Compared with the controls, a lower pH24 value was observed (P = 0.05) in ST muscle of carnitine-supplemented pigs, which – in castrates only – was associated with an increased drip loss (P < 0.01). Meat quality traits in m. longissimus were not influenced by treatment. In conclusion, our hypothesis that early-postnatal carnitine supplementation to piglets of low birth weight permanently increases myofibre number and improves later carcass and meat quality could not be confirmed by this experiment.

Diergeneeskundig gebruik van cardiogene biomarkers : focus op het gebruik bij paarden

Cardiale biomarkers, zoals natriuretische peptiden en troponinen, worden reeds routinematig in de humane geneeskunde gebruikt voor de diagnose, prognose en monitoring van hartaandoeningen. Hoewel ze ook in de kleine huisdierensector regelmatig bepaald worden om cardiologische en niet-cardiologische aandoeningen van elkaar te onderscheiden, is de literatuur omtrent het gebruik bij paarden beperkt. Bij paarden werd reeds een significante correlatie aangetoond tussen het ‘atrial natriuretic peptide’ (ANP), N-terminal ANP (NT-proANP) en de voorkamergrootte. Specifieke equine testen voor ‘brain natriuretic peptide’ (BNP) of ‘N-terminal BNP’ (NT-proBNP) bepaling ontbreken echter nog steeds. Troponinen (vooral troponine I) zijn meer uitgebreid bestudeerd bij het paard. Ze worden reeds gebruikt voor de diagnose van myocardschade en vervangen de minder specifieke lactaatdehydrogenase en creatinekinase iso-enzymen, waardoor het gebruik van deze laatste niet meer gerechtvaardigd is. Een laatste mogelijk belangrijke biomarker is aldosteron, maar aangezien slechts één studie bij paarden een correlatie aangetoond heeft tussen de aldosteronconcentratie en hartaandoeningen is uitgebreider onderzoek nodig.

1101 Are Lactat Dehydrogenase and Neuron Specific Enolase Analyses Good Diagnostic Tools for Assessing Extension of Perinatal Hypoxic-Ischemic Brain Injury?

BackgroundDiagnosis of perinatal hypoxic-ischemic encephalopathy (HIE) and early prediction neurological outcome is important and difficult. The aim of this study is to determine the value of neuron specific enolase (NSE)...

Inhibition kinetics of Plasmodium Lactate Dehydrogenase with herbal extracts suggest possible enzyme inhibitor molecular interaction

Background Resistance acquired by Plasmodium species (especially P. falciparum )t o most of the present antimalarial drugs is the principal hindrance in controlling malaria. Thus, one of the major challenges towards elimination of Malaria is development of novel and sustainable antimalarial drugs. In this milieu, herbs traditionally used to treat malaria are promising repertoires of antimalarial drugs. Earlier, lab studies have reported selective inhibition of P. falciparum and P. vivax specific Lactate Dehydrogenase (PfLDH and PvLDH) by Phyllanthus amarus aqueous extract and Murraya koenigii chloroform extract respectively. In the present investigation, we studied inhibition kinetics of PfLDH and PvLDH to explore molecular interactions between enzyme and inhibitor. Materials and methods Recombinant PfLDH and PvLDH, expressed in E. coli, were used in enzyme assay. LDH activity was measured in the direction of pyruvate to L-lactate conversion [1]. Steady state kinetic constants for substrate and cofactor as well as inhibition constants for plant extracts were measured by double reciprocal plot (Lineweaver and Burk plot). The enzyme inhibitor interactions were determined based on variations in the kinetic constants in presence of inhibitors, compared to control. Results Enzyme inhibition kinetics results are summarised in Table 1. Conclusion

Ethnopharmacology guided screening of traditional Indian herbs for selective inhibition of Plasmodium specific lactate dehydrogenase

Ethnopharmacological relevanceMedicinal plants traditionally used to treat malaria can provide quality leads towards identifying novel anti-malarial drugs. Here we combined this approach with target based drug discovery and explored Plasmodium specific lactate dehydrogenase (LDH) inhibitory activity of 8 Indian plants which are ethnically used to treat malaria. MethodsLDH from Indian Plasmodium falciparum and Plasmodium vivax strains, were cloned and expressed in Escherichia coli, followed by purification of recombinant enzymes (rPfLDH and rPvLDH respectively). Extracts of 8 plants in different organic and aqueous solvents, were screened for their inhibitory activity on rPfLDH, rPvLDH and mammalian LDHs. Phyllanthus amarus aqueous extract was further tested for in vitro parasiticidal activity. ResultsAqueous extract of Phyllanthus amarus Schum. and Thonn. and chloroform extract of Murraya koenigii (L.) Spreng. exhibited profound and exclusive inhibitory effect on Plasmodium falciparum LDH (IC50=11.2μg/ml±0.4) and Plasmodium vivax LDH (IC50=6.0μg/ml±0.6) respectively. Moreover, Phyllanthus amarus aqueous extract also demonstrated antiplasmodial activity in vitro, on Chloroquine sensitive and resistant strains of Plasmodium falciparum (IC50=7.1μg/ml±0.5 and 6.9μg/ml±0.7 respectively). ConclusionTarget specific screening of traditional herbs used in malaria treatment has proffered Phyllanthus amarus and Murraya koenigii extracts as hits which can optimistically provide novel antimalarial drugs.

Evaluation of the rapid diagnostic test CareStart pLDH Malaria (Pf-pLDH/pan-pLDH) for the diagnosis of malaria in a reference setting

BackgroundThe present study evaluated CareStart pLDH Malaria, a three-band rapid diagnostic test detecting Plasmodium falciparum-specific parasite lactate dehydrogenase (Pf-pLDH) and pan Plasmodium-specific pLDH (pan-pLDH) in a reference setting.MethodsCareStart pLDH was retrospectively and prospectively assessed with a panel of stored (n = 498) and fresh (n = 77) blood samples obtained in international travelers suspected of malaria. Both panels comprised all four Plasmodium species; the retrospective panel comprised also Plasmodium negative samples. The reference method was microscopy corrected by PCR. The prospective panel was run side-to-side with OptiMAL (Pf-pLDH/pan-pLDH) and SDFK60 (histidine-rich protein-2 (HRP-2)/pan-pLDH).ResultsIn the retrospective evaluation, overall sensitivity for P. falciparum samples (n = 247) was 94.7%, reaching 98.7% for parasite densities > 1,000/μl. Most false negative results occurred among samples with pure gametocytaemia (2/12, 16.7%) and at parasite densities ≤ 100/μl (7/12, 58.3%). None of the Plasmodium negative samples (n = 96) showed visible test lines. Sensitivities for Plasmodium vivax (n = 70), Plasmodium ovale (n = 69) and Plasmodium malariae (n = 16) were 74.3%, 31.9% and 25.0% respectively. Wrong species identification occurred in 10 (2.5%) samples and was mainly due to P. vivax samples reacting with the Pf-pLDH test line. Overall, Pf-pLDH test lines showed higher line intensities compared to the pan-pLDH lines (67.9% and 23.0% medium and strong line intensities for P. falciparum). In the prospective panel (77 Plasmodium-positive samples), CareStart pLDH showed higher sensitivities for P. falciparum compared to OptiMAL (p = 0.008), lower sensitivities for P. falciparum as compare to SDFK60 (although not reaching statistical significance, p = 0.08) and higher sensitivities for P. ovale compared to both OptiMAL (p = 0.03) and SDFK60 (p = 0.01). Inter-observer and test reproducibility were good to excellent.ConclusionCareStart pLDH performed excellent for the detection of P. falciparum, well for P. vivax, but poor for P. ovale and P. malariae.

Survival in metastatic castration resistant prostate cancer (mCRPC) trial participants.

e15136 Background: Median overall survival (mOS) in patients (pts) with metastatic prostate cancer progressing despite castrate levels of testosterone (mCRPC) was 13-16 months (m) in the pre-docetaxel era. These data, obtained from clinical trials, were used to construct currently available prognostic nomograms. We hypothesise that these models no longer reflect survival. Pts and physicians urgently require updated prognostic data on which to base management decisions. Methods: Pts with mCRPC treated on phase I-III trials at our institution were identified and data retrospectively collected. Predicted survival by Halabi and Smaletz nomograms were compared to calculated survival using Kaplan-Meier analysis. Cox model multivariate (MV) analysis used variables at referral, including performance status (PS), Gleason (GS), prostate specific antigen (PSA), lactate dehydrogenase (LDH), alkaline phosphatase (ALP), hemoglobin (Hb), visceral disease and albumin. Results: 423 pts with CRPC treated between 2003 and 2011 were included. At diagnosis median age was 62 years (y; 41.8 – 82.7); 226 (53.4%) had metastatic disease. Median interval from diagnosis to CRPC was 2.7y (0.2 – 21.7). At referral 248 pts (58.6%) were chemotherapy-naïve. Halabi and Smaletz models predicted mOS in chemo-naïve pts of 21m and 18m respectively, however the observed mOS was 32m (95%CI 28 – 38). Survival from CRPC was 43m (CI 37 – 46) and 39m (CI 34 - 44) in pre- and post-chemo pts, respectively. Conclusions: Despite aggressive disease characteristics, our pts lived significantly longer than predicted by current nomograms. MV analysis confirmed the importance of several previously identified prognostic factors. Survival data from this large cohort of CRPC pts should encourage men considering clinical trial participation. Previously developed nomograms no longer accurately predict survival.