Lactate Dehydrogenase Isoenzymes Research Articles

The study on the influence of blood specimen collection to the hospital inspection results

Objective To investigate the blood specimen collection site, inspection time and hemolysis influence on biochemical test results. Methods The specimens of 2586 cases outpatients and hospitalized patients in our hospital who accepted blood routine and blood coagulation four check were analyzed.The hemolysis and qualified specimens of biochemical test results were compared. Results 2 586 specimens found unqualified blood samples in 18 cases, and the incidence rate was 0.70% (18/2 586). Samples of different section and source were compared and found there were no significant difference (F=0.36, P>0.05). In 18 cases of unqualified blood samples due to technical problems and unqualified sample accounted for 88.89% (16 / 18). The technical factors that lead to unqualified samples was significantly higher than that of the instrument factor or fixed factors, and the difference was statistically significant(χ2=152.63, 98.52, all P<0.05). In addition, hemolysis caused the most of the unqualified samples for the technical factors.To choose potassium infusion on the side ipsilateral to the site or from the opposite side of the site collected blood samples.Sodium, chlorine, creatinine, uric acid, glucose detection found in effect of blood on the same side of the infusion of the biochemical test results in varying degrees.Potassium, chlorine, sodium, creatinine decreased significantly, and uric acid, glucose and other indicators were significantly increased (t=4.51, 4.98, 5.50, 3.25, 10.18, 15.25, all P<0.05). Gamma transpeptidase, creatine kinase, sodium and uric acid and other index of hemolytic samples were significantly lower than those of normal samples, and alanine aminotransferase (ALT), direct bilirubin, potassium, phosphorus, serum creatinine and creatine kinase were significantly higher than those of the normal samples (t=14.85, 21.21, 7.15, 4.86, 10.33, 4.02, 3.11, 8.20, 7.54, 5.11, all P<0.05). This may cause the depth in erythrocytes and some related substances.The time of collecting blood to sent to the room had obvious influence on the biochemical test results, alanine aminotransferase, aspartate amino transferase, glucose, creatine kinase and hydroxybutyrate and project them 1 h after submission were significantly lower than those of the standard inspection, and 1 hour after the submission of lactate dehydrogenase and creatine kinase isoenzyme index were significantly higher than those of the standardize inspection means (t=2.95, 4.82, 2.88, 5.44, 4.05, 3.98, 4.89, all P<0.05). Conclusion Sampling sites, hemolysis and inspection time had effect on blood specimen, so for the collection of blood samples must be strictly in accordance with the requirements of the operation. Key words: Blood specimen; Collection; Biochemical test

Osthole ameliorates acute myocardial infarction in rats by decreasing the expression of inflammatory-related cytokines, diminishing MMP-2 expression and activating p-ERK.

Osthole, the active constituent of Cnidium monnieri extracts, has been shown to have a diverse range of pharmacological properties. In the present study, we aimed to evaluate the cardioprotective effects of osthole in a rat model of acute myocardial infarction (AMI). The rats with AMI were treated with 1, 3 and 10 mg/kg of osthole or the vehicle for 4 weeks. The infarct size of the rats with AMI was measured, and casein kinase (CK), the MB isoenzyme of creatine kinase (CK-MB), lactate dehydrogenase (LDH) and cardiac troponin T (cTnT) activities in the rats with AMI were analyzed using commercially available kits. The nuclear factor-κB (NF-κB), tumor necrosis factor‑α (TNF-α), interleukin (IL)-1β and IL-6 levels in whole blood from rats with AMI were also detected using commercially available kits. The levels of Toll-like receptors 2/4 (TLR2/4) and nucleotide-binding oligomerization domain-containing protein 1/2 (NOD1/2) were also detected by RT-qPCR. Moreover, the protein expression levels of endothelial nitric oxide synthase (eNOS) and mitogen-activated protein kinase (MAPK) cascades, including extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK) and p38, cyclooxygenase-2 (COX-2), as well as matrix metalloproteinase-2 (MMP-2) were all assayed by western blot analysis. Our results revealed that osthole markedly reduced the infarct size, and the levels of CK, CK-MB, LDH and cTnT in the rats with AMI, and that these cardioprotective effects may be associated with the inhibition of inflammatory reactions, the reduction in MMP-2 activity and the activation of MAPK cascades.

Monitoring a 'metabolic shift' after surgical resection of oral squamous cell carcinomas by serum lactate dehydrogenase.

Monitoring surgical removal of oral squamous cell carcinomas (OSCC) is being routinely performed through clinical and imaging follow-up. We analyzed the potential use of serum lactate, lactate dehydrogenase (LDH), and LDH isoenzymes (LDH 1-5) as biomarkers in blood for the monitoring of surgical removal of OSCC. Serum lactate, LDH, and LDH isoenzymes (LDH 1-5) have been prospectively assessed in healthy individuals (n = 19), patients with OSCC (n = 34: primary OSCC, n = 32 and recurrent OSCC, n = 2) before surgery and after curative tumor resection (n = 26). LDHA and LDHB expressions were analyzed by immunohistochemistry (IHC) in the same OSCC tumor specimen. Positive LDHA tumor tissue expression measured by IHC (n = 34/34, 100%) was significantly associated with increased serum LDH-5 (n = 24/34, 71%, P = 0.0258) but weak significantly associated with increased total serum LDH (n = 23/34, 68%, P = 0.0592). Positive LDHB tumor tissue expression measured by IHC (n = 25/34, 74%) was significantly associated with increased total serum LDH (P = 0.0172). After surgery, serum LDH and LDH-5 isoenzyme significantly decreased and LDH-1 significantly increased in the aftercare. A significantly inverse correlation of LDHA with LDHB IHC scores was found (P < 0.0001). The association of LDHA and LDHB measured by IHC with serum LDH indicates that analyzing this enzyme could serve as a favorable 'liquid biopsy' (non-invasive diagnostic tool) for OSCC.

Effects of Daily Astaxanthin and L-Carnitine Supplementation for Exercise-Induced Muscle Damage in Training Thoroughbred Horses

The effects of dietary supplementation with astaxanthin and L-carnitine on serum markers and clinical incidence rate of exercise-induced muscle damage were studied in training horses. Sixty-three healthy Thoroughbred horses were randomly assigned to two groups and received the same base diet and exercise training throughout this study. The supplement (supp.) group (n = 31) received daily supplementation with astaxanthin (75 mg) and L-carnitine (3,000 mg) for 8 weeks, and the control (cont.) group (n = 32) received no supplementation. Blood samples were collected after high-intensity exercise training at 5 weeks before supplementation, 3 days (3d), and 8 weeks (8w) after the start of the supplementation. The blood samples were analyzed for creatine kinase (CK) activity and lactate dehydrogenase isoenzyme-5 (LDH-5), and a retrospective study was carried out by analyzing medical records for symptoms of exercise-induced muscle damage in both groups. Five horses (two of supp. group and three of cont. group) were excluded from this study for other disease. In the cont. group, CK activity at 8w was significantly increased compared with 3d, whereas the supp. group showed no significant change. After 8w, the CK activity of the supp. group was significantly lower than that of the cont. group. Additionally, the LDH-5 concentration of the supp. group also tended to be lower as compared with that of the cont. group. The incidence rate of exercise-induced muscle damage was significantly lower in the supp. group compared with the cont. group. These results suggest that continuous dietary administration of astaxanthin and L-carnitine attenuates exercise-induced muscle damage in horses.

Implications of oxidative stress in occupational exposure to lead on a cellular level

The aim of this study was to determine oxidative alterations leading to cellular dysfunctions in Pb-exposed subjects by evaluating damage to all major classes of biomolecules in the cell, lipid peroxidation, protein and DNA damage and determine relationships between parameters of Pb toxicity and specific biomarkers of oxidative damage.Analysis was conducted of smelter workers with high blood Pb and urine aminolevulinic acid levels and slightly elevated values of coproporphyrin and erythrocyte protoporphyrin IX. Significant decreases of thiol groups and increases in carbonyl groups as protein degradation end products, and of nitrite were detected. Elevated rates of lipid peroxidation and rises in the activities of the antioxidant enzymes Cu–Zn superoxide dismutase and catalase were also observed. Both enzymes showed positive correlations with the blood lead levels and urine coproporphyrin, while thiol groups correlated negatively with the same indices. The genotoxic potential of lead was manifested through an increased number of DNA-damaged cells. Increased activities of serum lactate dehydrogenase isoenzymes indicated cellular damage in the lungs, kidneys, and liver. These lead-induced impairments should be taken into consideration in the assessment of Pb-related health hazards.

Characteristics of membrane-bound pool of leukocyte lactate dehydrogenase in healthy men of different age

The study of isoenzymes of leucocyte lactate dehydrogenase (LDH) in healthy men of different age showed a decrease in the membrane-bound pool of LDH-1 and LDH-4 in 40-year-old males as compared with 20-year-old males. The results indicate an age-related reduction of the reserve membrane-bound pool of several LDH isoforms.

Lactate dehydrogenase isoenzyme 3 and hemolysis in sickle cell anemia: a possible correlation?

To the editor: The concept that lactate dehydrogenase (LDH) is a useful clinical marker of hemolysis in sickle cell anemia (SCA) is still a subject of debate.[1][1],[2][2] Some previous data suggested that the increase in serum LDH in these patients was mainly due to tissue damage rather than

Paeoniflorin ameliorates acute myocardial infarction of rats by inhibiting inflammation and inducible nitric oxide synthase signaling pathways.

Paeoniflorin (PF) is the main active component of the commonly used Traditional Chinese Medicine peony, Paeonia Suffruticosa. PF has diverse biological functions and exhibits anti‑oxidative, anti‑inflammatory and anti‑apoptotic activity. Inducible nitric oxide synthase (iNOS) is a catalyzing enzyme that is involved in the synthesis of nitric oxide (NO). NO has an important regulatory role in the cardiovascular, immune and nervous systems. PF has previously been demonstrated to inhibit the gene expression of iNOS. The present study aimed to identify a potentially novel cytoprotective function of PF, and to elucidate its effects against myocardial ischemic damage in a rat model of acute myocardial infarction (AMI). PF was able to significantly decrease the myocardial infarct size as well as the activities of creatine kinase (CK), the MB isoenzyme of CK, lactate dehydrogenase and cardiac troponin T. In addition, in the PF‑treated groups, the expression levels of tumor necrosis factor‑α, interleukin (IL)‑1β, IL‑6 and nuclear factor‑κB were markedly inhibited. Furthermore, treatment with PF inhibited the activities and protein expression levels of iNOS. Decreased caspase‑3 and caspase‑9 activities were also observed in the AMI rat model treated with various doses of PF. The results of the present study indicated that the cardioprotective effects of PF may be associated with the inhibition of inflammation and iNOS signaling pathways.

Determination of optimal storage temperature and duration for analysis of total and isoenzyme lactate dehydrogenase activities in canine serum and cerebrospinal fluid.

Lactate dehydrogenase (LDH) exists as 5 isoenzymes in both serum and cerebrospinal fluid (CSF). Human studies have demonstrated that changes in LDH activity can be correlated with a particular disease. Conflicting reports regarding the stability of LDH made it necessary to determine storage conditions before further study of the diagnostic power of this enzyme's activity can be pursued in dogs. The purpose of this study was to optimize measurement of LDH activity and analysis of its isoenzyme profile in canine serum and CSF through proper storage. Serum and CSF were collected from 5 healthy dogs. Samples were stored at 22°C, 4°C, or -20°C for up to 2months. Total LDH activity was measured spectrophotometrically. Isoenzyme profiles were determined using the QuickGel LDH Isoenzyme technique and densitometric scanning. Retention of >70% LDH activity in stored samples was considered clinically acceptable. Serum and CSF stored at -20°C retained >85% of the total LDH activity for 4weeks, although CSF total LDH activity degraded by >10% within 24hours of storage. All serum LDH isoenzymes retained >85% activity for up to 4weeks at -20°C. CSF LDH isoenzyme activity degraded rapidly, therefore CSF LDH should be evaluated within 72hours to assure >75% of LDH isoenzyme activity. Proper storage at -20°C can optimize detection of total LDH activity and the LDH isoenzyme profile in canine serum and CSF. This information is important for evaluating the potential usefulness of LDH in veterinary medicine.

Lactate dehydrogenase as a possible indicator of reproductive capacity of boars

The aim of this study was to determine the activity of lactate dehydrogenase (LDH) and its isoenzymes in the blood serum and seminal plasma of boars from the breeding and insemination centre with regard to their age. Significant quantitative differences in the presence of isoenzymes between blood serum and seminal plasma were recorded. Isoenzyme LDH-C4 in the seminal plasma of breeding boars of both groups was identified in the position between the third and fourth fraction of LDH, although boars from the breeding centre showed significantly lower concentrations. Refrigeration had a negative influence on the activity of LDH isoenzymes in the blood serum after 7 days and in seminal plasma after 3 days of storage. The work extends the knowledge in an insufficiently examined field of veterinary medicine: the distribution and physiological function of LDH in boars focusing on isoenzyme LDH-C4 in seminal plasma.

INMAP overexpression inhibits cell proliferation, induces genomic instability and functions through p53/p21 pathways.

INMAP is a spindle protein that plays essential role for mitosis, by ensuring spindle and centromere integrality. The aim of this study was to investigate the relevant functions of INMAP for genomic stability and its functional pathway. We overexpressed INMAP in HeLa cells, resulting in growth inhibition in monolayer cell cultures, anchorage-independent growth in soft agar and xenograft growth in nude mice. In this system caused micronuclei (MNi) formation, chromosome distortion and γH2AX expression upregulation, suggesting DNA damage induction and genomic stability impairment. As a tumour biochemical marker, lactate dehydrogenase (LDH) isoenzymes were detected to evaluate cell metabolic activity, the results confirming that total activity of LDH, as well as that of its LDH5 isoform, is significantly decreased in INMAP-overexpressing HeLa cells. The levels of p53 and p21 were upregulated, and however, that of PCNA and Bcl-2, downregulated. Indirect immunofluorescence (IIF) and coimmunoprecipitation (CoIP) analyses revealed the interaction between INMAP and p21. These results suggest that INMAP might function through p53/p21 pathways.

Comparison of multidetector computed tomography findings with clinical and laboratory data in pulmonary thromboembolism.

SummaryBackgroundPulmonary thromboembolism (PTE) is a common disease with a high mortality rate that is difficult to diagnose and treat. Because of the variety of clinical symptoms and signs, it is difficult to diagnose. Therefore, the diagnosis of PTE is mainly confirmed by imaging techniques. The aim of this study was to evaluate whether there is any corelation of the Wells rule, D-dimer and LDH values with computerized tomography pulmonary angiography (CTPA) findings in PTE diagnosis.Material/MethodsA consecutive series of 62 patients, which included 31 males and 31 females, with high/moderate/low risk of embolism according to Wells pulmonary embolism score, selected from the emergency service and/or outpatient clinic, enrolled in this prospective study. The patients with clinical or laboratory findings of elevated D-dimer level or elevated lactate dehydrogenase (LDH) level were suspected of embolism and underwent tomography.ResultsPTE was detected in 26 patients (42%). A significant difference was not detected between tomography finding positive and negative embolisms in the patient group in terms of age or gender distribution (P=0.221 and P=0.416, respectively).No significant difference was detected between tomography finding positive and negative embolisms in the patient group in terms of elevated LDH or/and D-dimer levels (P=0.263 and P=1.000, respectively).The distribution of low-risk-factor patients in the non-embolism group, and the distribution of high-risk-factor patients in the embolism-positive group was statistically significantly high (P<0.001). There was no statistically significant difference between the groups (P=0.053).Correlation test showed no correlation between LDH and D-dimer levels. (r=0.214, P=0.180).ConclusionsIn conclusion, when a patient presents with chest pain, our carrying out LDH and D-Dimer tests will not exclude PTE without CTPA. However, we suggest that LDH isoenzymes should be studied in further research.

Infection of sheep with Strongyloides papillosus: The effect of intensity of parasitic infection as well as therapy with albendazole on certain biochemical parameters in the blood of sheep

The objective of this work was to investigate certain biochemical parameters in the blood of sheep in conditions of different intensity of parasitic infection with Strongyloides papillosus, and also after the therapy with albendazole. The research was carried out on the sheep of Wurtemberg breed (n = 30) in which there was determined low, medium and high intensity of parasitic infection with S. papillosus. The control group (n = 10) consisted of the sheep that were negative to the presence of the parasitic infection. The degree and type of changes were monitored by determining the concentration of glucose, total protein, albumin, Albumin/Globulin ratio, AST, urea, total bilirubin, calcium, phosphorus, lactate dehydrogenase (LDH) total activity and isoenzyme distribution LDH1-5. On the basis of the obtained results it was concluded, through isoenzyme distribution of lactate dehydrogenase, that parasitic infection with S. papillosus can cause damage to the liver, heart muscle and lungs, while after the therapy with albendazole, the most affected organ is the liver. The concentration of glucose, total protein and albumin linearly decreased with the increase of intensity of the parasitic infection (p &lt; 0,05), and after the therapy with albendazole, a drop in concentration of these parameters was at a statistically significantly higher level (p&lt; 0,01). AST activity as well as the concentration of urea and total bilirubin grew linearly along with the intensity of the parasitic infection (p &lt; 0,05). After the therapy with ABZ, AST activity and concentration of total bilirubin were at a statistically significantly higher level (p &lt; 0,001), while urea concentration kept the same level as in the case of the parasitic infection. The values of calcium concentration (p &gt; 0,05) as well as phosphorus (p &lt; 0,05) linearly decreased with the parasitic infection intensity growth. Trend towards the decline of the concentration of these macroelements continues even after the therapy with albendazole (p &lt; 0,001). Considering our previous research in the field of oxidative stress, a phenomenon that is fundamentally related to the changes determined in this work, we strongly recommend that besides antihelmintics, preparations with antioxidative properties should be used in antiparasitic protocols as well. &lt;br&gt;&lt;font color="red"&gt;&lt;b&gt; This article has been retracted. Link to the retraction &lt;u&gt;&lt;a href="http://dx.doi.org/10.2298/VETGL190125001E"&gt;10.2298/VETGL190125001E&lt;/a&gt;&lt;u&gt;&lt;/b&gt;&lt;/font&gt;

Inter-Specific Biochemical Diversity between &amp;lt;i&amp;gt;Echis pyramidum&amp;lt;/i&amp;gt; and &amp;lt;i&amp;gt;Eryx colubrinus&amp;lt;/i&amp;gt; Inhabiting El-Faiyum, Egypt

Discontinuous polyacrylamide gel electrophoreses for lactate dehydrogenase (Ldh) and alfa-esterase (α-Est) isoenzymes were conducted for biochemical differentiation between Echis pyramidum and Eryx colubrinus inhabiting El-Faiyum, Egypt. Total lipids and total protein of liver and muscle tissues in both species were also analyzed. Three Ldh isoforms were recorded in both species and the activity and relative front (RF) of Ldh-1 seemed to be higher in E. pyramidum than in E. colubrinus. This high activity could be supported by the significant increase in the total lipids and total protein in liver and muscle tissues of this species. It is thus possibly reasonable to suppose that E. pyramidum is more active, energetic and adaptable in its habitat than E. colubrinus. α-Est showed five isozymic forms fractions in E. pyramidum, while it showed only four isoforms in E. colubrinus. α-Est-1 was the first clear, dense and thick isoform in E. pyramidum, but it was completely absent in E. colubrinus. High activity of the esterase isoform; α-Est-1 only in the heart tissue of E. pyramidum may reflect the high ability of E. pyramidum to be more resistant to the accumulated toxic residues in its body tissues than E. colubrinus.

Impairment of bone growth of wistar rat fetuses of diabetic and hypercholesterolemic mothers

Patients with either diabetes or hypercholesterolemia develop atherosclerosis and impair healing of bone. In the present work we illustrated their role in limb ischemia during bone cells differentiation of Wistar rat fetuses. Pregnant Wistar rats (n = 20 each) were arranged in three groups: control, diabetic or hypercholesterolemic. Diabetes was induced at the fifth day of gestation using streptozotocin, and hypercholesterolemia was carried by feeding virgin rats a diet containing 3% cholesterol for 6 wks prior to the onset of conception. At 13, 15, 17, and 19 d prenatal, pregnant rats were sacrificed, dissected, and fetuses were removed. Hind limbs were separated and subjected to histological and transmission electron microscopic examination, ossification, total calcium content of fetuses, isoenzymes alkaline and acid phosphatase and lactic dehydrogenase electrophoresis and DNA damage. Fetuses of diabetic or hypercholesterolemic mothers exhibited delayed histo-cytological differentiation of chondrocytes, and decreased periosteal ossification. Alkaline and acid phosphatase as well as lactic dehydrogenase isoenzymes showed altered diffusion rate and intensities of their bands reflecting their activities in both diseases comparing with the control. Assessments of bone calcium contents revealed marked reduction. Genomic expression of the degree of laddering (total DNA fragmented) or single-cell gel electrophoresis was found to be increased in cartilage and bone cells of fetuses of diabetic or hypercholesterolemic mothers. The authors concluded that both diseases had a selective, dramatic effect during fetus development in this model by retarding the histo- and cytological differentiation during limb bone growth. Both diseases increased the average cell death in skeletal elements and blood vessels as a consequence of altered alkaline and acid phosphatases and lactic dehydrogenase isoenzymes in accordance with DNA damage.

Oxysophoridine attenuates the injury caused by acute myocardial infarction in rats through anti-oxidative, anti-inflammatory and anti-apoptotic pathways.

Oxysophoridine (OSR), a natural alkaloid derived from the traditional Chinese medicinal plant sophora alopecuroides, can perform a variety of pharmacological actions. The aim of the present study was to assess the cardioprotective effect of OSR against acute myocardial infarction (AMI) in rats. OSR markedly reduced infarction size and levels of specific myocardial enzymes, including creatine kinase, the MB isoenzyme of creatine kinase, lactate dehydrogenase and cardiac troponin T. A reduced level of malondialdehyde was observed, and elevated catalase, Cu/Zn-superoxide dismutase (SOD), Mn-SOD, non-enzymatic scavenger glutathione and glutathione peroxidase activity were also identified in the OSR-treated rats. Additionally, OSR inhibited the activities of various inflammatory cytokines in a dose-dependent manner. These included nuclear factor-κB p65, tumor necrosis factor-α, and interleukin-1β, -6 and -10. Furthermore, OSR treatment suppressed caspase-3 activity in a dose-dependent manner. These results demonstrate that OSR ameliorates cardiac damage in a rat model of AMI and that this cardioprotection may be linked with its anti-oxidative, anti-apoptotic and anti-inflammatory properties.

Preventive effects of zingerone on altered lipid peroxides and nonenzymatic antioxidants in the circulation of isoproterenol-induced myocardial infarcted rats.

The present study was designed to evaluate the preventive effects of zingerone on circulatory lipid peroxides and nonenzymatic antioxidants in isoproterenol-induced myocardial infarcted rats. Rats were pretreated with zingerone (6 mg/kg body weight) daily for a period of 14 days and were then induced myocardial infarction with isoproterenol (100 mg/kg body weight) on 15th and 16th day. Increased intensities of serum lactate dehydrogenase isoenzymes 1 and 2 bands enhanced plasma lipid peroxidation products and lowered nonenzymatic antioxidant system were noted in isoproterenol-induced rats. Pretreatment with zingerone daily for 14 days revealed significant preventive effects on the electrophoretic and biochemical parameters evaluated in isoproterenol-induced rats. Furthermore, the in vitro study confirmed the potent antioxidant activity of zingerone. The results of our study showed that zingerone protected the rat's heart against isoproterenol-induced myocardial infarction by its antioxidant effect.

Search of substances with the cardioprotective activity in the series of derivatives of halogenides 1-alkyl- and carboxyalkyl-4-ylideneamino-1,2,4-triazole

In spite of achievements in the sphere of highly effective drugs creation for treating cardiovascular diseases the problem is still urgent since mortality is high because of these diseases, and it takes the 2-nd – 3-d place among the population of industrially developed countries. In the Research and Production Association “Pharmatron” a new approach concerning creation of new effective anti-anginal drugs has been developed; it is a chemical modification of position 1 and 4 of 1,2,4-triazole molecule by introduction of the structural fragments of the most active medicines and structures imitating sites of adrenergic receptors. Bromides of 1-(alkyl) carboxyalkyl-4-ylideneamino-1,2,4-triazole have been synthesized at the Pharmaceutical Chemistry department of Zaporizhzhia State Medical University under the supervision of professor I.A.Mazur. The cardioprotective activity of 12 compounds – derivatives of 1-carboxyalkyl-4-ylideneamino-1,2,4-triazole bromides and 1-alkyl-4-ylidenamino-1,2,4-triazole bromides has been investigated. The experiments were conducted in 510 white outbred male rats weighing 120-130 g from the mouse bank of the Institute of Pharmacology and Toxicology of the AMS of Ukraine. Determination of acute toxicity of the compounds investigated was conducted according to Kerber method. It has been found that LD50 of these compounds when introduced intraabdominally to rats is within 29.5-295 mg/kg. It allows to refer the compounds under research to the III and IV classes of toxicity (moderate or low toxic). The model of acute myocardial infarction in rats was used for studying the cardioprotective activity of the compounds, the infarction was modeled by staged introduction of isadrin and pituitrin. It has been found that intraabdominal introduction of 8 compounds of 12 ones in the dose of 1/100 LD50 to rats with myocardial infarction leads to decrease of hyperenzymemia of cardiospecific isoenzymes of creatine phosphokinase (MB CPK) and lactate dehydrogenase (LDH-1), which play the role of biochemical markers of the myocardium damage, and decrease of electrophysiological marker ΣΔST during electrocardiography. The structural fragments of the molecule, which play a determinative role in manifestation of the cardioprotective effect of such series as β-phenylethyl, carboxy propyl, octyl in position 1 and the presence of amino- and n-methoxybensilidenamino-groups in position 4, have been revealed. It has been determined that compound MT significantly exceeds the therapeutical efficacy of metoprolol by decrease of MB CPK and LDH-1 activity in the blood serum of rats with myocardial infarction, as well as decrease of ECG ST amplitude.

Lactate dehydrogenase (LD), alkaline phosphatase (ALP) isoenzymatic patterns in Iraqi children with visceral leishmaniasis before and after treatment with stibogluconate.

The mean levels of alkaline phosphatase (ALP), lactate dehydrogenase enzymes exhibited a significant elevation in visceral leishmaniasis (VL) patients compared to the control. There was no significant change in relation to the sex and age. ALP isoenzymes revealed three banding patterns which differ from the three zymodems which were obtained from control group. These differences may be due to isoenzymes activity of patients with VL before and after therapy. Lactate dehydrogenase (LD) isoenzymes revealed five banding patterns differ from the five normal zymodems. These differences mainly occurred due to LD isoenzymes activity in patients with VL before and after therapy.

ISOENZYME ELECTROPHORESIS AND KARYOTYPE TECHNIQUES AS AN ALTERNATIVE FOR CELL LINE IDENTITY CONFIRMATION

The use of cell culture in biomedical science has been constantly enhanced and the use of certified and microrganisms-free cell lines is indispensable. Due to the value and the difficulty of acquiring a commercially available kit (AuthentikitTM system) for our routine in addition with the purpose of monitoring and ensuring quality of provided cell lines, and also contributing with cell characterization, the Núcleo de Cultura de Células do Instituto Adolfo Lutz (NCC– AL) padronized two techniques to identify the animal species from which the cells were originated: isoenzyme electrophoresis and karyotype. Cell extracts from 14 different cell lines were run under horizontal electrophoresis in agarose gel and lactate dehydrogenase isoenzyme profile was revealed using tetrazolium salt. The number of bands and the migration distances between bands of cell lines were similar to the literature, but small adjustments were needed to assist in the visualization of the bands that were not visible. Cell flasks for karyotype were incubated with colcemid solution and modal values of each cell line were calculated. Comparing all the results obtained with those reported in the literature, we conclude that both methods are promising and will allow better control of the services provided by the laboratory, not to mention guaranteeing authenticated cell lines for research and diagnostics. DOI: http://dx.doi.org/10.17525/vrr.v19i1.90