Total LDH Activity Research Articles

Betanin protects against bleomycin-induced pulmonary fibrosis by regulating the NLRP3/IL-1β/TGF-β1 pathway-mediated epithelial-to-mesenchymal transition.

Idiopathic pulmonary fibrosis (IPF) is a life-threatening disease that leads to dyspnea and progressive loss of lung function. This study aimed to investigate the protective effect of betanin (BET), the major pigment in red beetroot, on pulmonary fibrosis induced by bleomycin (BLM) in rats and to assess the underlying mechanisms. In this view, total and differential cell counts and LDH activity in bronchoalveolar lavage fluid were estimated. Furthermore, MDA and GSH contents in the lungs were colorimetrically measured, while hydroxyproline, NLRP3, ASC, caspase-1, TGF-β1, and vimentin levels in lung tissue were evaluated using the ELISA technique. Moreover, IL-1β, E-cadherin, and α-SMA expressions were analyzed by immunostaining of lung specimens. BET treatment protects against pulmonary fibrosis as indicated by the reduction in total and differential cell counts, LDH activity, hydroxyproline, NLRP3, ASC, caspase-1, IL-1β, and TGF-β1 levels. MDA content was also decreased following BET administration, while GSH content was elevated. Additionally, BET suppressed the EMT process as evidenced by an increase in E-cadherin expression besides the reduction in vimentin and α-SMA expressions. To conclude, these results revealed the protective effect of BET against pulmonary fibrosis that might be attributed to the attenuation of the NLRP3/IL-1β/TGF-β1 signaling pathway and EMT process.

The evaluation of reactive oxygen species generation and free mitochondrial DNA in platelet concentrates during 5 days of storage.

Oxidative stress and mitochondrial damage are causes of platelet storage lesions (PSLs). Mitochondrial damage causes mitochondrial DNA (mtDNA) to be released into the extracellular space. MtDNA in platelet concentrates is considered damage-associated molecular patterns (DAMPs) and is one of the major causes of PSLs. The mechanism of mtDNA release in platelet concentrates has not been thoroughly investigated. This study aimed to determine the effect of reactive oxygen species (ROS) on mtDNA release in platelet concentrates during storage. Ten platelet concentrates from healthy donors were obtained in this investigation. Platelet concentrates were prepared by platelet-rich plasma (PRP) and stored at 22 ± 2 C° with gentle agitation. Platelet concentrates were subjected to flow cytometry and real-time PCR to evaluate total ROS and free mtDNA on days 0, 3, and 5 of platelet concentrate storage. Total ROS detected significantly increased from day 0 to day 5 of platelet concentrate storage (P = 0.0079). The mean of copy numbers of free mtDNA on day 0 increased from 3.43 × 106 ± 1.57 × 106 to 2.85 × 107 ± 1.51 × 107 (molecules/μl) on the fifth day of platelet concentrate storage, and it was statistically significant (P = 0.0039). In addition, LDH enzyme activity significantly increased during platelet concentrate storage (P < 0.0001). Also, releasing mtDNA in platelet concentrates was directly correlated with total ROS generation (P = 0.021, r = 0.61) and LDH activity (P = 0.04, r = 0.44). The evidence from this study confirmed the increasing level mtDNA copy numbers in platelet concentrates during storage, and the amount of free mtDNA is directly correlated with ROS generation and platelet lysis during 5 days of platelet concentrate storage. Finally, these changes may be related to DAMPs in the platelet concentrates.

SPECIES- AND AGE-SPECIFIC CHARACTERISTICS OF THE LACTATE DEHYDROGENASE SYSTEM IN TISSUES OF RODENTS (MAMMALIA: RODENTIA)

The aim of this study was to analyze the species-specific features and age-related changes in the total lactate dehydrogenase activity (LDh, EC 1.1.1.27) and LDh patterns in some tissues in three species of rodents (Rodentia) with different ecological characteristics – the northern birch mouse Sicista betulina (Pallas, 1779), the European water vole Arvicola amphibius (L., 1758) and the tundra vole Microtus oeconomus (Pallas, 1776). Compared to the European water vole and the northern birch mouse, the tundra vole had a higher content of M-subunits and the amount of LDh-5 isoenzyme in the liver and skeletal muscles, higher contents of aerobic LDh-1 isoenzymes and, respectively, of h-subunits in kidneys and heart. In the heart and kidneys, the hybrid fractions (LDh-2, LDh-3, LDh-4) dominated over LDh-1 and LDh-5 in the northern birch mouse. The total LDh activity in the liver and kidneys of the northern birch mouse was higher than in the tundra vole. The extent of anaerobic ATP production in the liver of small rodents varied – the liver of the diving European water vole contained more LDh-2, LDh- 3, LDh-4 and h-subunits than the corresponding tissue in the non-diving tundra vole. In kidneys of the northern birch mouse and European water vole, the content of the LDh- 5 isoenzyme increased through the ontogeny. The LDh activity was higher in the heart of European water vole adults compared to young animals. Species-specific differences in LDh isoenzyme spectra and total LDh activity in the studied mammals reflect their adaptation to the semi-aquatic and hibernating lifestyles.

Granulomatous lung inflammation is nanoparticle type-dependent

ABSTRACTBackground: Nanoparticles are increasingly suspected as a strong etiologic factor of granuloma formation. Aim of the Study: The aim of our study was to compare lung inflammatory response and histology changes following exposure of mice to two widely used nanoparticles: carbon nanotubes (MWCNT) and cadmium-based nanoparticles (QDOT705) in an attempt to better our understanding of granulomatous inflammation. Materials and Methods: Various groups of mice were included: control mice and mice that were intranasally instilled with QDOT or MWCNT. At defined time points post-challenge, bronchoalveolar lavages (BALs) and lung tissues were collected to study inflammatory and histologic changes. Results: Analyses of lung BAL fluids and tissues of nanoparticles-challenged mice in comparison to controls found: (1) increased cellularity in BALs, (2) increase of total protein concentration, LDH activity and proteolytic activity in BALs; (3) patchy granulomas, (4) macrophages, CD3 ± T, Treg and B cell infiltration in granulomatous areas; and (5) altered regulation of key inflammatory mediators and receptors. Importantly, these changes were nanoparticle type-dependent. Conclusion: Our work enhances understanding of nanoparticles-induced lung inflammatory and histological changes that result in granuloma formation. We provide compelling evidence that not only exposure to nanoparticles leads to granulomatous lung inflammation, but the severity of this latter is nanostructure type-dependent. Of importance, while nanotechnology has the potential to revolutionize various fields including medicine, nanoparticles form the potential for an entirely new lung health risk that it is necessary to take seriously into consideration by setting up and/or reinforcing adequate safety measures.

Lanthanum chloride reduces lactate production in primary culture rat cortical astrocytes and suppresses primary co-culture rat cortical astrocyte-neuron lactate transport.

Lanthanum (La) can impair learning memory and induce behavioral abnormalities in animals. However, the mechanism underlying these adverse effects of La is still elusive. It has been demonstrated that lactate derived from astrocytes is the major energy source for neurons during long-term memory (LTM) formation and the deficiency of lactate supply can result in LTM damage. However, little work has been done with respect to the impact of La on the lactate production in astrocytes and astrocyte-neuron lactate transport (ANLT). Herein, experiments were undertaken to explore if there was such an adverse effect of La. Primary culture rat cortical astrocytes and primary co-culture rat cortical astrocyte-neuron were treated with (0.125, 0.25 and 0.5mM) lanthanum chloride (LaCl3) for 24h. The results showed that LaCl3 treatment significantly downregulated the mRNA and protein expression of glucose transporter 1 (GLUT1), glycogen synthase (GS), glycogen phosphorylase (GP), lactate dehydrogenase A (LDHA), and monocarboxylate transporter 1, 2 and 4 (MCT 1 2 and 4); upregulated the mRNA and protein expression of lactate dehydrogenase B (LDHB); and decreased the glycogen level, total LDH and GP activity, GS/p-GS ratio and lactate contents. Moreover, rolipram (20, 40μM) or forskolin (20, 40μM) could increase the lactate content by upregulating GP expression and the GS/p-GS ratio, as well as antagonize the effects of La. These results suggested that La-induced learning-memory damage was probably related to its suppression of lactate production in astrocytes and ANLT. This study provides some novel clues for clarifying the mechanism underlying the neurotoxicity of La.

Lanthanum damages learning and memory and suppresses astrocyte-neuron lactate shuttle in rat hippocampus.

Rare-earth elements (REEs) are applied in various fields by virtue of their superior physical and chemical properties. Surveys have reported that REEs can impair learning and memory in children and induce neurobehavioral abnormalities in animals. However, the mechanism underlying this neurotoxicity is still unclear. Lanthanum (La) is often chosen to study the effects of REEs. Here, we investigated the role of astrocyte-neuron lactate shuttle (ANLS) in spatial learning and memory impairment induced by LaCl3 in hippocampus, an important spatial memory-related brain region. Pregnant Wistar rats were exposed to 0, 0.125, 0.25, 0.5, or 1% LaCl3 in drinking water during pregnancy and lactation. After weaning, young rats continued to receive 0, 0.125, 0.25, 0.5, and 1% LaCl3 in the drinking water for 1month. The results showed that LaCl3 exposure impaired the spatial learning and memory of rats in Morris water maze test, significantly reduced the mRNA and protein levels of glycogen synthetase, glycogen phosphorylase, lactate dehydrogenase A, monocarboxylate transporter 4, MCT-1, and MCT-2, and decreased total LDH activity and lactate contents in rat hippocampus. These results indicate that LaCl3 impairs spatial learning and memory in rats probably by suppressing ANLS in rat hippocampus. The study provides a novel clue of energy supply for neurons to clarify the neurotoxicity of REEs.

The expression of Ldh-c in the skeletal muscle of plateau pika (Ochotona curzoniae) enhances adaptation to a hypoxic environment.

ABSTRACTThe plateau pika (Ochotona curzoniae) is a species of sprint-running alpine animals in the Qinghai-Tibet Plateau, which is a harsh highland hypoxic environment. Ldh-c is expressed in the testis, sperm and somatic tissues of plateau pika. To reveal the role and physiological mechanisms of sperm-specific lactate dehydrogenase (LDH-C4), in plateau pika to adapt to hypoxic environment, an adenoviral line of pMultiRNAi-Ldhc was constructed and injected into the bilateral biceps femoris of the hind legs. The swimming times of the pikas, and the Ldh-c expression levels, total LDH activities and ATP levels in skeletal muscle, were measured after the pikas were raised in the trapped site for 5 days. Our results showed that after Ldh-c was silenced, the sprint-running ability (swimming time) of the plateau pikas was significant decreased, and the total LDH activities and ATP levels were reduced by 28.21% and 27.88%, respectively. Our results indicated that expression of Ldh-c in the skeletal muscle of plateau pika increased anaerobic glycolysis and enhanced adaptation to highland hypoxic environments.

In Vivo Enzymes Activities of Some Ru(II) Compounds with N-Alkylphenothiazines

Abstract The purpose of the present study was to investigate and compare the effects of two ruthenium complexes with trifluoperazine on acethylcholinesterase enzyme activity and lactate dehydrogenase levels in vivo under physiological conditions in rats blood. Complexes 1 and 2 showed positive effects on acethylcholinesterase at all doses and did not disturb its normal activity. Total LDH activity was inhibited in the presence of both complexes, but Ru(II) complexes showed different effects on the activity of LDH isoenzymes. The activities of LDH1 and LDH2 isoenzymes were decreased in all applied doses of the complex 2, while the activity of LDH2 reduced using complex 1 in the same doses. Results of the present study suggest the neuro- and cardio protective potential of oral administration of complexes 1 and 2, as non-toxic compounds under physiological conditions. These protective effects are the result of their potent antioxidant activity.

The Effects of Ferulic Acid Against Oxidative Stress and Inflammation in Formaldehyde-Induced Hepatotoxicity.

This study was designed to elucidate the protective effects of ferulic acid (FA) on formaldehyde-induced hepatotoxicity by measuring some routine biochemical parameters, cytokine levels, and oxidative stress-related parameters in addition to YKL-40 in male Wistar albino rats. Tissue superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px) activities, and tissue malondialdehyde (MDA) levels were measured. Also, serum YKL-40, TNF-α, IL-6, IL-1β, IL-8, total protein, albumin, total bilirubin concentrations, and AST, ALT, ALP, and LDH activities were measured. Histological specimens were examined in light microscopy. Formaldehyde significantly increased tissue MDA, and serum cytokine levels and also decreased activities of antioxidant enzymes. FA treatment decreased MDA and cytokine levels and increased activities of antioxidant enzymes. FA also alleviated degeneration due to formaldehyde toxicity. We suggested that FA can be used as a promising hepatoprotective agent against formaldehyde toxicity because of the obvious beneficial effects on oxidative stress parameters.

Abstract 15099: Suppression of Autophagic Flux by Sequestration of p62 Promotes RIP1-mediated Necroptosis in Cardiomyocytes

Purpose: Necroptosis, a programmed form of necrosis, has been implicated in various pathological conditions including heart failure. Here we examined whether and how receptor-interacting protein kinase (RIP) 1-dependent necroptosis is functionally linked with autophagy in cardiomyocytes. Methods and results: In H9c2 cells, the necroptotic pathway was activated by TNF-α (TNF, 50 ng/ml) alone or by co-incubation of TNF with 20 mM z-VAD-fmk (zVAD), a pan-caspase inhibitor. TNF increased LDH activity in the culture medium from 26.6±4.3% of total LDH activity in the vehicle-treated control to 39.1±4.8%, suggesting induction of necrosis. Addition of zVAD to TNF (TNF/zVAD) further increased the LDH activity to 70.6±2.7%. The augmented cell death by TNF/zVAD was completely reversed by necrostatin-1 (39.4±4.0%), a RIP1 inhibitor, and was attenuated by rapamycin (10 nM), a promoter of autophagy (49.4±2.0%). Immunoblot analyses showed that TNF/zVAD significantly increased the level of LC3-II. The accumulation of LC3-II protein was not further increased by bafilomycin A1 (100 nM), an inhibitor of lysosomal degradation, indicating that accumulation of LC3-II by TNF/zVAD reflected suppression of autophagic flux. Immunoprecipitation experiments revealed that TNF/zVAD reduced interaction between LC3-II and p62, an autophagy adaptor protein, and conversely increased interaction between RIP1 and p62. The changes in p62 interactosomes by TNF/zVAD were reversed by pretreatment with rapamycin. On the other hand, TNF/zVAD-induced cell death when p62 was knocked down was still sensitive to necrostatin-1, indicating that a p62-independent pathway is also involved in RIP1-mediated necroptosis. Conclusions: Suppression of autophagic flux by p62 sequesteration from p62-LC3-II complexes and its recruitment to RIP1 promotes RIP1-dependent necroptosis by TNF/zVAD.

Determination of hematology and plasma chemistry reference intervals for 3 populations of captive Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus).

The imperiled status of Atlantic sturgeon (Acipenser oxyrinchus oxyrinchus), a large, long-lived, anadromous fish found along the Atlantic coast of North America, has prompted efforts at captive propagation for research and stock enhancement. The purpose of this study was to establish hematology and plasma chemistry reference intervals of captive Atlantic sturgeon maintained under different culture conditions. Blood specimens were collected from a total of 119 fish at 3 hatcheries: Lamar, PA (n=36, ages 10-14years); Chalk Point, MD (n=40, siblings of Lamar); and Horn Point, Cambridge, MD (n=43, mixed population from Chesapeake Bay). Reference intervals (using robust techniques), median, mean, and standard deviations were determined for WBC, RBC, thrombocytes, PCV, HGB, MCV, MCH, MCHC, and absolute counts for lymphocytes (L), neutrophils (N), monocytes, and eosinophils. Chemistry analytes included concentrations of total proteins, albumin, glucose, urea, calcium, phosphate, sodium, potassium, chloride, and globulins, AST, CK, and LDH activities, and osmolality. Mean concentrations of total proteins, albumin, and glucose were at or below the analytic range. Statistical comparisons showed significant differences among hatcheries for each remaining plasma chemistry analyte and for PCV, RBC, MCHC, MCH, eosinophil and monocyte counts, and N:L ratio throughout all 3 groups. Therefore, reference intervals were calculated separately for each population. Reference intervals for fish maintained under differing conditions should be established per population.

Evaluation of the Physiological Relationship Between Oxidative Stress and Metabolic Characters: Insights From Hybridization Between Anarhichas minor × A. lupus

Physiological indicators such as lipid peroxidation and enzymes involved in aerobic/anaerobic energy metabolism were measured in pure and hybrid lines of wolffish juveniles (Anarhichas minor, ♀ A. minor ⨯ ♀ A. lupus). Interspecific hybridization was introduced as a powerful tool to promote augmentation of phenotypic variability and consequently improve our capacity for the detection of clear physiological relationships amongst oxidative stress and metabolic characters. Mitochondrial sequence genes analysis between the Atlantic (A. lupus) and spotted wolffish (A. minor) were later performed to further assess the extent of divergence between A. lupus and A. minor mitochondrial haplotypes. The observed divergences (2.0% and the number of nonsynonymous substitution n=23) do not seem to be sufficient to generate detectable differences in ETS complexes catalytic capacities. Total LDH activity was however higher in hybrids than in the pure strands and tightly correlated to TBARS levels which suggest that LDH actively participates to the regulation of cellular redox status. Our results are discussed in comparison with a similar study on a charr hybrid (Salvelinus alpinus × S. fontinalis).

Anthocyanin determination in blueberry extracts from various cultivars and their antiproliferative and apoptotic properties in B16-F10 metastatic murine melanoma cells

Blueberry consumption is associated with health benefits contributing to a reduced risk for cardiovascular disease, diabetes and cancer. The aim of this study was to determine the anthocyanin profile of blueberry extracts and to evaluate their effects on B16-F10 metastatic melanoma murine cells. Seven blueberry cultivars cultivated in Romania were used. The blueberry extracts were purified over an Amberlite XAD-7 resin and a Sephadex LH-20 column, in order to obtain the anthocyanin rich fractions (ARF). The antioxidant activity of the ARF of all cultivars was evaluated by ABTS, CUPRAC and ORAC assays. High performance liquid chromatography followed by electrospray ionization mass spectrometry (HPLC-ESI-MS) was used to identify and quantify individual anthocyanins. The anthocyanin content of tested cultivars ranged from 101.88 to 195.01 mg malvidin-3-glucoside/100g fresh weight. The anthocyanin rich-fraction obtained from cultivar Torro (ARF-T) was shown to have the highest anthocyanin content and antioxidant activity, and inhibited B16-F10 melanoma murine cells proliferation at concentrations higher than 500 μg/ml. In addition, ARF-T stimulated apoptosis and increased total LDH activity in metastatic B16-F10 melanoma murine cells. These results indicate that the anthocyanins from blueberry cultivar could be used as a chemopreventive or adjuvant treatment for metastasis control.

The pulmonary toxicity study of nano-silica particles on rats through dynamic inhalation

To investigate the pulmonary toxicity of different concentrations of nano-silica (nano-SiO2) under continuous dynamic inhalation conditions in the rat. 48 male Sprague-Dawley rats were randomly divided into four groups, including the dispersant control group (saline) and nano-SiO2 low-dose group (0.3%, w/v), the middle-dose group (1%) and the high-dose group (3%). Animals were sacrificed at 28 d after exposure under continuous dynamic inhalation conditions, and bronchoalveolar lavage fluid (BALF) and lung tissue were collected. And following items were observed: body coefficient, BALF related items (leukocytes and classification, total protein content, LDH activity), lung tissue pathological changes (HE staining), and pulmonary fibrosis forming (collagen fiber VG staining). Compared to the dispersant control group, there was no significant change on lung organ coefficient in Nano-SiO2 group (P < 0.05). The BALF total WBC count in 1% and 3% in nano-SiO2 groups showed higher value than the dispersant control group (P < 0.05). No obvious changes were found on total protein content and LDH activity in nano-SiO2 groups compared to the dispersant control group (P > 0.05). For differential WBC counts, lymphocyte count in BALF in nano-SiO2 groups was significantly decreased (P < 0.05), monocyte and macrophage counts were significantly increased (P < 0.05), but there was no difference on the proportion of neutrophils (P > 0.05). HE staining results showed that the obvious thickening of alveolar wall in nano-SiO2 groups, inflammatory cell infiltration also increased around the bronchial and vascular wall. Lung fibrosis VG staining showed no significant change of collagen fiber distribution. Under our experimental conditions, the continuous dynamic inhalation of nano-SiO2 only caused the significant inflammation in rat lungs, pulmonary fibrosis phenomenon could not be observed significantly.

The effect of the intensity of parasitic infection with Strongyloides papillosus and albendazole therapy on biochemical parameters in sheep blood

The aim of this report was to study the biochemical parameters in sheep blood under conditions of various intensities of parasitic infection with Strongyloides papillosus, as well as after therapy with albendazole (ABZ). Investigations were performed on sheep of the Wurtemberg race (n = 30) in which were detected mild, moderate and high intensities of parasitic infection with S. papillosus. The control group (n = 10) was composed of sheep negative to parasitic infections. The degree and type of changes were monitored by determining the concentrations of glucose, total proteins, albumin, A/G ratio, AST, urea, total bilirubin, calcium, phosphorus, total LDH activity and isoenzymatic LDH1-5 distributions. On the basis of the obtained results, we determined, through isoenzymatic LDH distribution, that during parasitic infection with S. papillosus, there is ongoing damage to the liver, heart muscle and lung, while after therapy with ABZ, the liver suffers the most damage. The concentration of glucose, total proteins and albumin fell linearly with the rise in the intensity of parasitic infection (p<0.05), while after therapy with ABZ, the fall of these parameters was at a statistically significantly higher level (p<0.01). The activity of AST, concentration of urea and total bilirubin also rose linearly with the intensity of parasitic infection (p<0.05). After therapy with ABZ, the activity of AST and the concentration of total bilirubin were at a statistically significantly higher level (p<0.001), while the concentration of urea retained the same levels as in the case of parasitic infection. Values of calcium concentrations (p>0.05) and phosphorus (p<0.05) also fall linearly with the rise of the intesity of the parasitic infection. The trend in the concentration fall of these macroelements, continues also after treatment with albendazole (p<0.001). Having in mind our previous studies in the field of oxidative stress and phenomena lying behind these changes, we strongly recommend that in antiparasitic treatment protocols, beside antihelminthics, compounds with antioxidative properties should also be used.

Evaluation of CdSe/CdS-PEG-FA quantum dots: distribution and observable-adverse-effect-level in mice after intravenous injection

Tissue distribution and observable-adverse-effect-level of cadmium selenide/cadmium sulfide (CdSe/CdS) quantum dots (QDs) were investigated to get important information of this QDs. Female BALB/c mice were treated with single intravenous (IV) injection of polyethylene glycol-folic acid-coated QDs (CdSe/CdS-PEG-FA) at different concentrations (0, 270, and 540 pmol/20 g of mice body weight), and the subsequent toxicological effect-levels were examined for 24 h. The health and apparent behaviors of the animals were normal throughout the study. The distribution of the QDs was observed in the spleen, liver, lung and kidneys, but not in the brain and heart tissues. The spleen and liver possess the highest amount of the QDs followed by the lung while the kidneys possess the few. There were no changes in the organ weight index, total protein concentration, LDH activity, and specific NADPH oxidase activity in any tested organ indicating no toxic effects of the QDs in our study. Additionally, histopathological examination did not show any cellular/tissue structural damages. Overall, single IV administration of CdSe/CdS-PEG-FA QDs to BALB/c mice allows immediate systematic availability, and showed different tissue distribution without any obvious toxic effects at our experimental design.

Biomarkers of acute respiratory allergen exposure: Screening for sensitization potential

Effective hazard screening will require the development of high-throughput or in vitro assays for the identification of potential sensitizers. The goal of this preliminary study was to identify potential biomarkers that differentiate the response to allergens vs non-allergens following an acute exposure in naïve individuals. Female BALB/c mice received a single intratracheal aspiration exposure to Metarhizium anisopliae crude antigen (MACA) or bovine serum albumin (BSA) in Hank's Balanced Salt Solution (HBSS) or HBSS alone. Mice were terminated after 1, 3, 6, 12, 18 and 24 h. Bronchoalveolar lavage fluid (BALF) was evaluated to determine total and differential cellularity, total protein concentration and LDH activity. RNA was isolated from lung tissue for microarray analysis and qRT-PCR. MACA administration induced a rapid increase in BALF neutrophils, lymphocytes, eosinophils and total protein compared to BSA or HBSS. Microarray analysis demonstrated differential expression of genes involved in cytokine production, signaling, inflammatory cell recruitment, adhesion and activation in 3 and 12 h MACA-treated samples compared to BSA or HBSS. Further analyses allowed identification of ∼ 100 candidate biomarker genes. Eleven genes were selected for further assessment by qRT-PCR. Of these, 6 demonstrated persistently increased expression (Ccl17, Ccl22, Ccl7, Cxcl10, Cxcl2, Saa1), while C3ar1 increased from 6–24 h. In conclusion, a single respiratory exposure of mice to an allergenic mold extract induces an inflammatory response which is distinct in phenotype and gene transcription from the response to a control protein. Further validation of these biomarkers with additional allergens and irritants is needed. These biomarkers may facilitate improvements in screening methods.

Protective Effect of L-Carnitine and Coenzyme Q10 on CCl4-Induced Liver Injury in Rats

This study provides an information about the mechanisms of liver injury induced by CCl4, and determines the influence of administration of L-carnitine or/and CoQ10 as prophylactic agents against CCl4 deteriorative effect. The study was carried out on 80 adult male albino rats divided into eight groups, 10 animals each, as follows: four normal groups (control, treated with L-carnitine, treated with CoQ10, and treated with a combination of Lcarnitine and CoQ10) and four liver injury groups treated with CCl4 (control, treated with L-carnitine, treated with CoQ10, and treated with a combination of L-carnitine and CoQ10). Liver injury was induced by s.c. injection of a single dose of CCl4 (1 ml/kg). L-carnitine (50 mg/kg/day) was given i.p. for four successive days 24 hours before CCl4 injection, and CoQ10 (200 mg/kg) was given as a single i.p. dose 24 hours before CCl4 injection. Animals were sacrificed 24 hours after CCl4 injection, blood samples were withdrawn and liver tissue samples were homogenized. The levels of the following parameters were determined: hepatic reduced glutathione, serum ALT and AST, hepatic lipid peroxides, hepatic vitamin C, hepatic and serum total protein, serum albumin, serum sialic acid, serum nitrite, and serum and hepatic total LDH activities and LDH isoenzymes. The obtained data revealed that CCl4 injection produced a significant decrease in reduced glutathione content, vitamin C, total protein and albumin levels. However, there was a significant increase in serum ALT and AST activities, lipid peroxides, sialic acid, nitric oxide, serum and hepatic total LDH activities. On the other hand, groups treated with L-carnitine or/and CoQ10 prior to CCl4 injection showed an improvement in most parameters when compared with cirrhotic control group. It has been concluded that L-carnitine and coenzyme Q10 have a pronounced prophylactic effect against liver damage induced by halogenated alkanes such as carbon tetrachloride.

Lactate dehydrogenase activity of rat epididymis and spermatozoa: effect of constant light.

During its passage through the epididymis, the gamete undergoes a process of "maturation" leading to the acquisition of its fertilizing ability. The epididymis displays regional variations in the morphology and metabolic properties of its epithelium which are relevant for the progressive development of mature sperm characteristics. The epididymis has spontaneous peristaltic contractions and receives sympathetic innervation that is modulated by melatonin, a hormone synthesized and released by the pineal gland. Constant lighting disrupts melatonin synthesis and secretion. We have studied the effect of constant light on lactate dehydrogenase (LDH; EC 1.1.1.27) and its isozyme C4 activities and protein content in whole epididymis, epididymal tissue and in spermatozoa from caput and cauda segments. Animals were exposed from birth to an illumination schedule of 14 h light:10 h dark (group L:D). At 60 days of age one group of animals was submitted to constant light over 50 days (group L:L). In order to test the fertilizing ability, the rats of each group were mated with soliciting estrous females. The percentage of pregnancies in females mated with males maintained in L:L was remarkably lower than those in females mated with males maintained in the L:D photoperiod (44% and 88% respectively). Constant light increased protein concentration and LDH activity in caput as well as in cauda of total epididymis. On the contrary, in epididymal tissue, the protein content decreased in both epididymal sections compared with controls. When enzymatic activity was expressed in Units per spermatozoa, constant light induced a significant reduction of total LDH and LDHC4 in caput and cauda spermatozoa while LDH activity of epididymal tissue was not affected. In spite of the decrease in LDH per sperm cell when rats were exposed to constant light, in total epididymis (epididymis tissue plus sperm cells content) and in spermatozoa, values of enzyme activities expressed per weight unit were higher than those of controls. This is explained by the increase in the amount of stored spermatozoa, both in caput and cauda, produced by exposure of animals to constant light. Our results confirm that in rats, chronic exposure to constant light promotes a reduction of fertilizing ability and indicates that continuous lighting reduces the total LDH and LDHC4 activities, possibly due to moderate aging of spermatozoa within the duct by lengthening of the sperm transit through the epididymis.

Neuroprotective effect of carvedilol and melatonin on 3-nitropropionic acid-induced neurotoxicity in neuroblastoma

In neurodegenerative diseases, progressive oxidative stress is a major event that precedes neuronal death. Oxidative stress is characterized by an imbalance between oxidants and antioxidants. This imbalance induced oxidative molecular and cell damage, reducing cellular viability. 3-Nitropropionic acid (3NP) causes oxidative stress and other molecular and cellular changes similar to those observed in neurons of patients with Huntington's disease. Since carvedilol and melatonin act as free-radical scavengers, this study examined the effect of carvedilol (10(-5) M) and melatonin (10(-5) M) on oxidative and cell damage induced by 3NP in N1E-115 neuroblastoma cells. Carvedilol and melatonin prevented the increases in lipid peroxidation and total LDH activity, as well as the depletion of reduced glutathione (GSH) and the reduction of antioxidative enzymes activities in N1E-115 cells incubated with 100 mM 3NP. All these carvedilol and melatonin effects were more intense when the drugs were added before rather than after inducing the damage by 3NP. These results also provided evidence supporting the hypothesis that carvedilol and melatonin can be useful for treating neurodegenerative diseases, such as Huntington's disease.