Increase In Tissue Content Research Articles

Effect of Cigarette Smoke Exposure and Aspirin Treatment on Neurotransmitters' Tissue Content in Rats' Hippocampus and Amygdala.

Cigarette smoke withdrawal can cause anxiety-like behavior and modulate neurotransmitter-related proteins in the brain. We examined the effects of cigarette smoke with and without aspirin treatment on the concentrations of neurotransmitters, including dopamine, serotonin, glutamate, glutamine, and GABA in the amygdala and hippocampus. Sprague-Dawley rats were randomly assigned to four different groups: (1) control group exposed only to standard room air, (2) cigarette smoke exposed group treated with saline vehicle, (3) cigarette smoke exposed group treated with aspirin (30 mg/kg), and (4) control group treated only with aspirin (30 mg/kg). Cigarette smoke exposure was performed for 2 h/day, 5 days/week, for 31 days. Behavioral testing was carried out weekly, 24 h after cigarette smoke exposure, during acute withdrawal. At the end of week 4, rats were given either distilled water (1 mL) or aspirin 45 min before cigarette exposure for 11 days. Dopamine, serotonin, glutamate, glutamine, and GABA were extracted from both the amygdala and hippocampus and were separated and quantified using a developed and validated HPLC-MS/MS method. Cigarette smoke withdrawal induced anxiety behaviors, and aspirin treatment reduced this effect. Cigarette smoke exposure increased tissue content of dopamine, serotonin, glutamate, glutamine, and GABA, and aspirin treatment reversed this effect. Cigarette smoke caused an increase in tissue content of several neurotransmitters as well as anxiety-like behavior, and these effects were normalized by aspirin treatment.

Impact of Dietary Palmitic Acid on Lipid Metabolism.

Palmitic acid (PA) is ubiquitously present in dietary fat guaranteeing an average intake of about 20 g/d. The relative high requirement and relative content in the human body, which accounts for 20–30% of total fatty acids (FAs), is justified by its relevant nutritional role. In particular physiological conditions, such as in the fetal stage or in the developing brain, the respectively inefficient placental and brain blood–barrier transfer of PA strongly induces its endogenous biosynthesis from glucose via de novo lipogenesis (DNL) to secure a tight homeostatic control of PA tissue concentration required to exert its multiple physiological activities. However, pathophysiological conditions (insulin resistance) are characterized by a sustained DNL in the liver and aimed at preventing the excess accumulation of glucose, which result in increased tissue content of PA and disrupted homeostatic control of its tissue concentration. This leads to an overaccumulation of tissue PA, which results in dyslipidemia, increased ectopic fat accumulation, and inflammatory tone via toll-like receptor 4. Any change in dietary saturated FAs (SFAs) usually reflects a complementary change in polyunsaturated FA (PUFA) intake. Since PUFA particularly n-3 highly PUFA, suppress lipogenic gene expression, their reduction in intake rather than excess of dietary SFA may promote endogenous PA production via DNL. Thereby, the increase in tissue PA and its deleterious consequences from dysregulated DNL can be mistakenly attributed to dietary intake of PA.

Anti-Hyperglycaemic and Anti-Oxidative Effect of Merremia Tridentata (Linn) Hallier F Ethanol Leaf Extract

The anti-hyperglycaemic and anti-oxidative effect of Merremia tridentata (Linn) Hallier F ethanol leaf extract (MTELE) was evaluated in this study. Diabetes was induced on male albino wistar rats by a single intra-peritoneal injection of 65mg/kg body weight dose of streptozotocin. The rats were divided into seven groups and treated accordingly with varying doses (50 mg/kg, 75 mg/kg and 100 mg/kg body weight) of MTELE for seven weeks. Body weight and plasma glucose levels were monitored while antioxidant parameters, catalase (CAT), superoxide dismutase (SOD) activities, reduced glutathione (GSH) concentration and levels of thiobarbituric acid reacting substances (TBARS) were determined in the liver and kidney tissues. There was a significant (p<0.05) reduction in the weight of the diabetic control group which was significantly (p<0.05) ameliorated by the treatment. Plasma glucose levels were significantly (p<0.05) reduced by 50.6%, 62.1% and 72.68% in diabetic rats treated with 50 mg/kg, 75 mg/kg and 100mg/kg body weight dose of MTELE respectively. There was a significant (p<0.05) improvement in all the antioxidant parameters investigated in this study in both the liver and kidney tissues at all levels of treatment (increase in the activities of CAT and SOD, increase in tissue content of GSH and drastic reduction of TBARS level). The extract at 100mg/kg body weight dose on normal rats showed a significant (p<0.05) improvement in the status of SOD in the kidney, GSH and TBARS in the liver when compared with the normal control group. MTELE demonstrated a good anti-hyperglycaemia and antioxidant potential and should be further investigated for development of lead compound capable of preventing and treating oxidative stress related complications in diabetics.

Low cardiac content of long-chain acylcarnitines in TMLHE knockout mice prevents ischaemia-reperfusion-induced mitochondrial and cardiac damage

Increased tissue content of long-chain acylcarnitines may induce mitochondrial and cardiac damage by stimulating ROS production. N6-trimethyllysine dioxygenase (TMLD) is the first enzyme in the carnitine/acylcarnitine biosynthesis pathway. Inactivation of the TMLHE gene (TMLHE KO) in mice is expected to limit long-chain acylcarnitine synthesis and thus induce a cardio- and mitochondria-protective phenotype. TMLHE gene deletion in male mice lowered acylcarnitine concentrations in blood and cardiac tissues by up to 85% and decreased fatty acid oxidation by 30% but did not affect muscle and heart function in mice. Metabolome profile analysis revealed increased levels of polyunsaturated fatty acids (PUFAs) and a global shift in fatty acid content from saturated to unsaturated lipids. In the risk area of ischemic hearts in TMLHE KO mouse, the OXPHOS-dependent respiration rate and OXPHOS coupling efficiency were fully preserved. Additionally, the decreased long-chain acylcarnitine synthesis rate in TMLHE KO mice prevented ischaemia-reperfusion-induced ROS production in cardiac mitochondria. This was associated with a 39% smaller infarct size in the TMLHE KO mice. The arrest of the acylcarnitine biosynthesis pathway in TMLHE KO mice prevents ischaemia-reperfusion-induced damage in cardiac mitochondria and decreases infarct size. These results confirm that the decreased accumulation of ROS-increasing fatty acid metabolism intermediates prevents mitochondrial and cardiac damage during ischaemia-reperfusion.

Effects of Garden Amendments on Soil Available Lead and Plant Uptake in a Contaminated Calcareous Soil

Gardeners use organic and inorganic substances to enhance plant growth, which can inadvertently impact soil solubility and plant uptake of unknown contaminants. Consequently, human exposure can increase through gardening and consumption of produce grown in potentially contaminated soils. A greenhouse experiment was established to examine the effects of biochar, compost, and common inorganic fertilizer on soil lead (Pb) availability for radish (Raphanus sativus, L.) and lettuce (Lactuca sativa, L.) grown in a calcareous soil containing excessively high lead (Pb), along with Pb accumulation in radish tissue. Results indicate that soil amended with biochar and planted to radish saw an 18% reduction in available Pb and an 11% decrease in plant tissue content when compared to the control. Compost showed an 8% reduction in available Pb, but a 19% increase in tissue content. In contrast, soil with inorganic fertilizer planted to radish increased in both soil Pb availability by 11% and Pb tissue content by 40%. Adding water-soluble inorganic fertilizers to contaminated calcareous soils without added organic matter enhances soil Pb availability and often asymptomatic plant Pb bioaccumulation. In conclusion, gardeners are encouraged to test their soils for contamination and apply biochar in combination with compost, as this combination is recommended to improve soil health and aid in overcoming initial N deficiencies induced by biochar.

CAN SUCRALFATE ENEMA PREVENT COLITIS IN COLONIC SEGMENTS WITHOUT FECAL TRANSIT?

ABSTRACT - BACKGROUND:Oxidative stress is one of the main mechanisms associated with the rupture of the defense mechanisms of the colonic epithelial barrier; it reduces the tissue content of the claudin-3 and occludin proteins, which are the main constituents of intercellular tight junctions. Sucralfate (SCF) has antioxidant activity and has been used to treat different forms of colitis.AIM: This study aimed to measure the tissue claudin-3 and occludin content of the colon mucosa without fecal transit, subjected to intervention with SCF.METHODS: Thirty-six rats were subjected to left colon colostomy and distal mucous fistula. They were divided into two groups according to euthanasia that was performed 2 or 4 weeks after the intervention. Each group was divided into three subgroups according to the enema applied daily: saline alone, SCF at 1 g/kg/day, or SCF at 2 g/kg/day. Colitis was diagnosed by the histological analysis adopting the previous validate scale. The tissue expression of both proteins was identified by immunohistochemical technique. The content of proteins was quantified by computer-assisted image analysis.RESULTS: The inflammatory score was high in colonic segments without fecal transit, and enemas with SCF reduced the inflammatory score in these segments, mainly in those animals submitted to intervention with SCF in greater concentration and for a longer period of intervention. There was an increase in tissue content of claudin-3 and occludin, related to SCF concentration. The tissue content of both proteins was not related to the intervention time.CONCLUSION: Enemas with SCF reduced the inflammation and increased the tissue content of claudin-3 and occludin in colonic mucosa without fecal stream.

Altered protein levels in bone marrow lesions of hip osteoarthritis: Analysis by proteomics and multiplex immunoassays

To assess tissue level changes of proteome and cytokine profiles of subchondral bone in hip osteoarthritis (OA) affected by bone marrow lesions (BMLs). We compared significant protein level differences in osteoarthritic bone with BMLs to control bone without bone marrow lesions. Subchondral bone biopsies were taken from femoral heads of end-stage osteoarthritis patients with (BML, n=21) and without (CON, n=9) BMLs. Proteins were extracted through a standardized Trizol protocol and used in the subsequent analyses. Angiogenesis and bone markers were assessed using multiplex immunoassays (Luminex). Liquid chromatography tandem mass spectrometry (LC-MS/MS) was performed to detect significant differences in proteome and peptide profiles between BML and CON. Multiplex immunoassays revealed increased tissue contents of vascular endothelial growth factors (VEGF-A/C/D), endothelin-1, angiopoietin-2 and interleukin-6 (IL-6) in bone with BMLs compared to control bone, whereas osteoprotegerin levels were reduced. Mass spectrometry demonstrated pronounced increase in the levels of hemoglobin (73-fold), serum albumin (30-fold), alpha-1-antitrypsin (9-fold), apolipoprotein A1 (4.7-fold), pre-laminin-A/C (3.7-fold) and collagen-alpha1-XII (3-fold) in BMLs, while aggrecan core protein (ACAN) and hyaluronan and proteoglycan link protein 1 (HAPL1) decreased 37- and 29-fold respectively. Reduced osteoprotegerin, ACAN and HAPL1 are consistent with osteoclastic activation and high remodeling activity in BMLs. The pronounced increase in angiogenesis markers, hemoglobin and serum albumin support the presence of increased vascularity in subchondral bone affected by BMLs in OA. VEGFs and IL-6 are known nociceptive modulators, and increased levels are in keeping with pain being a clinical feature frequently associated with BMLs.

Amelioration of Aflatoxin B1-induced gastrointestinal injuries by Eucalyptus oil in rats.

Background Eucalyptus oil (EO), derived from Eucalyptus species, possesses vast remedial and healing properties, although its gut health-promoting properties have not been well investigated. In this study, we investigated the chemical composition of a commercial EO formulation and its potential role in protecting against aflatoxin B1 (AfB1)-induced gastrointestinal damage in rats. Methods Male Wistar rats were divided into six groups with eight rats each. Control rats were administered with the vehicle (1% Tween 80) for 14 days, while another group was exposed to two oral doses of AFB1 on days 12 and 14. Two other groups were pre-treated with oral doses of EO (50 and 100 mg/kg b.w.) for 14 consecutive days, along with two oral doses of AfB1 (5 mg/kg b.w.) on days 12 and 14. The remaining two groups were treated with EO alone at the two doses for 14 days. At the end of the experiment, blood samples, stomach and intestinal tissues were collected for measurement of oxidative stress and antioxidant parameters and light microscopic examination. Results Gas chromatography-mass spectrometry analysis revealed Eucalyptol (1, 8-cineole) as the main constituent (67.48%) of the oil. AfB1 administration induced oxidative and inflammatory disturbances, indicated by significantly (p<0.05) increased serum nitric oxide level and myeloperoxidase activity; increased tissue contents of hydrogen peroxide, malondialdehyde and protein carbonyls, accompanied with corresponding histological alterations. AfB1 also induced significant (p<0.05) reductions in glutathione peroxidase and superoxide dismutase (SOD) activities. Treatment with EO produced significant improvements in the biochemical parameters as well as the appearance of the gastric and intestinal mucosa. EO alone, at the two doses tested did not produce any significant changes in the parameters investigated. Conclusion The findings from this study showed that EO demonstrated protective activity against Aflatoxin-induced toxicity in stomach and intestinal tissues and may thus find application in treatment of gastrointestinal disorders.

Promoted Growth and Development of Carnation Plantlets In Vitro by Ventilation and Combined Red and Blue Light

In this study, the principal objective was to investigate the effect of light quality and vessel ventilation on the growth and development, physiology, activities of antioxidant enzymes, and contents of mineral nutrients of carnation (Dianthus caryophyllus L.) ‘Marble Beauty’. Single node cuttings stuck into the plant growth regulator (PGR)-free MS medium in containers covered with caps with or without a ventilation filter were cultured first four weeks under white and then additional four weeks under either white (control), blue, red, or red + blue light emitting diodes (LEDs) for 56 days. Interestingly, a ventilated culture condition not only reduced the percentage of the hyperhydricity, but also increased the total chlorophyll content (Chl a + Chl b) of the plantlets as compared to the non-ventilated condition. In addition, blue LEDs produced plantlets with the greatest number of shoots and red LEDs produced plantlets with the greatest shoot length. The quality of plantlets was improved under a ventilation condition. Besides, under a ventilated condition, red + blue LEDs raised APX activity, and blue LEDs not only raised the activity of the CAT, but also increased tissue contents of such elements as K, Ca, Mg, Zn, Mn and Fe. The red LEDs increased contents of B and Si under a ventilated condition, and Na accumulation under a non-ventilated condition. Thus, including blue or red LEDs as the light source in a ventilated culture condition will produce plantlets of carnation ‘Marble Beauty’ in vitro with improved quality.

Effects of Increased Pharyngeal Tissue Mass Due to Fluid Accumulation in the Neck on the Acoustic Features of Snoring Sounds in Men.

Snoring sounds are generated by the vibration of pharyngeal tissue due to the upper airway narrowing. While recorded by a microphone placed over the neck, snoring can pass through the pharyngeal tissue surrounding the upper airway. Thus, changes in the pharyngeal tissue content may change the acoustic properties of the snoring sounds. Rostral fluid shift and the consequent increases in neck fluid volume (NFV) and neck circumference (NC) can increase pharyngeal tissue mass. Therefore, the goal of this study was to investigate the relationship between increases in pharyngeal tissue mass, as assessed by increased NFV and NC, and snoring sounds features. We obtained data from a previous study where 20 males who were not obese participated in a daytime polysomnography and their NC and NFV were measured before and after sleep. During sleep, snoring sounds were recorded with a microphone placed over the neck. Spectral centroid of the snoring sounds was estimated. Then, the first five snoring segments were selected from the first and last 30 minutes of stage N2 sleep. We found a significant decrease in the snoring spectral centroid from the beginning to end of sleep. We also found that spectral centroid from the end of sleep in frequency ranges below 200 Hz was inversely correlated with the increases in NFV and NC from before to after sleep. These results suggest that snoring spectral centroid can be used as a noninvasive and convenient method to assess variations in the pharyngeal tissue mass.

Palmitic Acid: Physiological Role, Metabolism and Nutritional Implications.

Palmitic acid (PA) has been for long time negatively depicted for its putative detrimental health effects, shadowing its multiple crucial physiological activities. PA is the most common saturated fatty acid accounting for 20–30% of total fatty acids in the human body and can be provided in the diet or synthesized endogenously via de novo lipogenesis (DNL). PA tissue content seems to be controlled around a well-defined concentration, and changes in its intake do not influence significantly its tissue concentration because the exogenous source is counterbalanced by PA endogenous biosynthesis. Particular physiopathological conditions and nutritional factors may strongly induce DNL, resulting in increased tissue content of PA and disrupted homeostatic control of its tissue concentration. The tight homeostatic control of PA tissue concentration is likely related to its fundamental physiological role to guarantee membrane physical properties but also to consent protein palmitoylation, palmitoylethanolamide (PEA) biosynthesis, and in the lung an efficient surfactant activity. In order to maintain membrane phospholipids (PL) balance may be crucial an optimal intake of PA in a certain ratio with unsaturated fatty acids, especially PUFAs of both n-6 and n-3 families. However, in presence of other factors such as positive energy balance, excessive intake of carbohydrates (in particular mono and disaccharides), and a sedentary lifestyle, the mechanisms to maintain a steady state of PA concentration may be disrupted leading to an over accumulation of tissue PA resulting in dyslipidemia, hyperglycemia, increased ectopic fat accumulation and increased inflammatory tone via toll-like receptor 4. It is therefore likely that the controversial data on the association of dietary PA with detrimental health effects, may be related to an excessive imbalance of dietary PA/PUFA ratio which, in certain physiopathological conditions, and in presence of an enhanced DNL, may further accelerate these deleterious effects.

Dopaminergic and behavioural changes in a loss-of-imprinting model of Cdkn1c.

The imprinted gene Cdkn1c is expressed exclusively from the maternally inherited allele as a consequences of epigenetic regulation. Cdkn1c exemplifies many of the functional characteristics of imprinted genes, playing a role in foetal growth and placental development. However, Cdkn1c also plays an important role in the brain, being key to the appropriate proliferation and differentiation of midbrain dopaminergic neurons. Using a transgenic model (Cdkn1c BACx1) with a twofold elevation in Cdkn1c expression that mimics loss‐of‐imprinting, we show that increased expression of Cdkn1c in the brain gives rise to neurobiological and behavioural changes indicative of a functionally altered dopaminergic system. Cdkn1c BACX1 mice displayed altered expression of dopamine system‐related genes, increased tyrosine hydroxylase (Th) staining and increased tissue content of dopamine in the striatum. In addition, Cdkn1c BACx1 animals were hypersensitive to amphetamine as showed by c‐fos expression in the nucleus accumbens. Cdkn1c BACX1 mice had significant changes in behaviours that are dependent on the mesolimbic dopaminergic system. Specifically, increased motivation for palatable food stuffs, as indexed on a progressive ratio task. In addition, Cdkn1c BACX1 mice displayed enhanced social dominance. These data show, for the first time, the consequence of elevated Cdkn1c expression on dopamine‐related behaviours highlighting the importance of correct dosage of this imprinted gene in the brain. This work has significant relevance for deepening our understanding of the epigenetic factors that can shape neurobiology and behaviour.

Fetal Brain Development Involves Activation of Inflammatory Pathways in the Fetal Cerebral Cortex: Increased Tissue Content of Interleukin 1β

Fetal Brain Development Involves Activation of Inflammatory Pathways in the Fetal Cerebral Cortex: Increased Tissue Content of Interleukin 1β

Peripheral nerve injury and gabapentin, but not their combination, impair attentional behavior via direct effects on noradrenergic signaling in the brain

Chronic pain after peripheral nerve damage is often accompanied by a reduction in prefrontal cortex (PFC)–related cognitive functions, which are regulated by noradrenaline, released from efferents originating in the locus coeruleus (LC). L5 to L6 spinal nerve ligation (SNL) in rats increased tissue content and extracellular concentrations of noradrenaline in microdialysates from the PFC, and impaired attentional level in the novel object recognition test. Systemic gabapentin, commonly used to treat chronic pain, impaired the novel object recognition task in normal but not SNL animals. Accordingly, gabapentin increased c-fos expression in LC neurons and noradrenaline release in the PFC in normal animals, but in SNL animals, gabapentin failed to increase c-fos expression in LC neurons projecting to the PFC and failed to increase noradrenaline release in the PFC. In contrast, locally perfused gabapentin reduced noradrenaline release in the PFC in vivo and in PFC synaptosomes in vitro. SNL- and gabapentin-induced impairment of novel object recognition task were reversed by intraperitoneal injection of the α1-adrenoceptor antagonist prazosin. These results suggest that increase in noradrenergic tone, induced by nerve injury or gabapentin, impairs PFC functions possibly via α1-adrenoceptor–mediated mechanisms; that the net effect of gabapentin on noradrenaline release in the PFC would depend on sometimes opposing actions at different sites; and that nerve injury selectively impairs the response to gabapentin in PFC-projecting neurons in the LC.

Impact of Ischemia-Reperfusion on Extracellular Matrix Processing and Structure of the Basement Membrane of the Heart

PurposeAcute ischemic injury is a strong inductor of cardiac remodelling, resulting in structural changes of the extracellular matrix (ECM) and basement membrane (BM). In a large animal model of ischemia-reperfusion (I/R) we investigated the post-ischemic liberation of the collagen-IV-fragments Tumstatin (TUM; 28 kDa-fragment of collagen-IV-alpha-3), Arresten (ARR; 26 kDa-fragment of collagen-IV-alpha-1) and Endorepellin (LG3, 85 kDa-fragment of perlecan) which are biologically active in angiogenesis and vascularization in the post-ischemic myocardium.Methods and ResultsIn this blinded study, 30 pigs were randomized to 60 min of global I/R at either 4°C or 32°C or served as control. Three transmyocardial tissue samples were collected prior to ischemia and within 30 min and 150 min of reperfusion. Tissue content of TUM, ARR and LG3 was analyzed by western blotting and immunostaining.Within 150 min of mild hypothermic I/R a significantly increased tissue content of ARR (0.17±0.14 vs. 0.56±0.56; p = 0.001) and LG3 (1.13±0.34 vs. 2.51±1.71, p<0.001) was observed. In contrast, deep hypothermic I/R was not associated with a significant release of cleavage products. Cleavage of TUM remained unchanged irrespective of temperature. Increased matrix processing following mild hypothermia I/R is further supported by a >11fold elevation of creatine kinase (2075±2595 U/l vs. 23248±6551 U/l; p<0.001) in the coronary sinus plasma samples. Immunostaining demonstrated no changes for ARR and LG3 presentation irrespective of temperature. In contrast, TUM significantly decreased in the BM surrounding cardiomyocytes and capillaries after mild and deep hypothermic I/R, thus representing structural alterations of the BM in these groups.ConclusionThe study demonstrates an early temperature-dependent processing of Col-IV as major component of the BM of cardiomyocytes and vascular endothelium. These observations support the protective effects of deep hypothermia during I/R. Furthermore, the results suggest an increased structural remodelling of the myocardial basement membrane with potential functional impairment during mild hypothermic I/R which may contribute to the progression to post-ischemic heart failure.

Effect of electrical stimulation of nucleus accumbens with low, median and high currents intensities on conditioned place preference induced by morphine in rats.

Background:Some investigators indicated the effect of electrical or chemical stimulation on different parts of the brain and its effect on animal's behaviors. Furthermore, drug addiction is known to be associated with dysfunction of memory and motivational systems. In this study, we aimed to evaluate the effect of electrical stimulation of nucleus accumbens (NAc) with different currents intensities on conditioned place preference (CPP) induced by morphine.Materials and Methods:Male Wistar rats were randomly divided for experimental groups (n = 8). We investigated the influence of electrical stimulation with different current intensities (low: 15 μA, median: 50 μA and high: 100 μA) on NAc with ineffective and effective dose of morphine (0.5 and 5 mg/kg, respectively) on acquisition and expression of morphine-induced place conditioning in male rats.Results:The doses of subcutaneous administration morphine (2.5 and 5 mg/kg, P < 0.05 and P < 0.001; respectively) induced CPP compared with saline group. Furthermore, our findings are showed that electrical stimulation (100 μA) of NAc suppressed morphine-induced CPP. It revealed impairment of learning and memory formation in conditioning process due to morphine administration.Conclusion:It is possible that high current intensity (100 μA) had an accompanied effect by a reversal of the increased tissue contents of dopamine and its metabolites in the NAc of morphine-induced CPP rats. Furthermore, high current intensity in combination with ineffective dose of morphine (0.5 mg/kg) increased morphine-induced CPP probability via the prove reward system.

Fish Omega-3 Fatty Acids Induce Liver Fibrosis in the Treatment of Bile Duct-Ligated Rats

Biliary atresia-induced cholestasis increases hepatic oxidative stress with eventual progression to cirrhosis and liver failure. Omega-3 fatty acids play a possible role in the regulation of oxidative stress and the improvement of cholestasis. The goal of the present study is to investigate the role of dietary supplementation of fish omega-3 fatty acids in the reduction of hepatocellular damage by using a rat common bile duct ligation model. Sprague-Dawley rats received either sham or bile duct ligation (BDL) and were divided into four study groups: Sham+saline (Sham+sal) group, Sham+Fish oil (Sham+FO) group, BDL+saline (BDL+sal) group, and BDL+Fish oil (BDL+FO) group. Rats from each group were assigned to receive, besides regular chow, once daily with either normal saline or fish omega-3 fatty acids (0.4 % of its own body weight) via gavage for 10 days. Samples of blood, liver tissue homogenates, and histological studies from different groups were analyzed at the end of the study. Rats from BDL+FO had significantly impaired liver function as compared to other study groups (p < 0.05 is of significant difference). Ishak scores and the TGF-b1 contents were significantly higher in rats that received BDL+FO, p < 0.05. Contrary to TGF-b1 liver content, rats from the BDL+FO group had the lowest glutathione levels among the study groups, p < 0.05. Fish omega-3 fatty acids supplementation, albeit increased tissue content of DHA, tended to increase liver fibrosis in BDL rats, decrease liver glutathione level, and compromise hepatic function; fish oil supplementation to subjects with biliary atresia might be of potential hazard and should be used with caution.

Searching for a therapy of creatine transporter deficiency: some effects of creatine ethyl ester in brain slices in vitro

Creatine, an ergogenic compound essential for brain function, is very hydrophilic and needs a transporter to cross lipid-rich cells' plasma membranes. Hereditary creatine transporter deficiency is a severe incurable neurological disease where creatine is missing from the brain. Creatine esters are more lipophylic than creatine and may not need the transporter to cross plasma membranes. Thus, they may represent a useful therapy for hereditary creatine transporter deficiency. Creatine ethyl ester (CEE) is commercially available and widely used as a nutritional supplement. It was reported that it enters the cells of patients lacking the transporter but was not useful when administered in vivo, by oral route, to affected patients. In this paper we investigated the effects of CEE in in vitro brain slices before and after biochemical block of the creatine transporter. We found that CEE is rapidly degraded in the aqueous incubation medium to creatinine, however it remains in solution long enough to cause an increase in tissue content of creatine and, more prominently, phosphocreatine. Both CEE and creatine delayed the anoxia-induced failure of synaptic transmission, and there was no difference between the two compounds. Contrary to what we expected, CEE did not increase tissue creatine content after the creatine transporter was blocked. We confirm that CEE is probably not an effective treatment for hereditary creatine transporter deficiency. Two factors seem to affect the possibility for creatine esters to be exploited in the therapy of creatine transporter deficiency. First, the size of their alcohol moiety should be increased since this would increase the lipophilicity of the compound and improve its ability to diffuse through biological membranes. Second, creatine esters should be further modified to slow their degradation to creatinine and increase their half-life in aqueous solutions. Moreover, we should not forget the possibility that they are degraded in vivo by plasma esterases.

Glycosaminoglycans in human normal lung tissues and lung cancer tissues

The quantitative changes of glycosaminoglycans in tumor tissue of human lung cancers (6 squamous cell carcinomas, 7 small cell carcinomas and 10 adenocarcinomas) were studied. Normal lung tissues contained of 3.38 mumol uronic acid/g dry weight glycosaminoglycans which consisted of hyaluronic acid, chondroitin sulfates, dermatan sulfate and heparan sulfate. The total amount of glycosaminoglycans in human lung cancer tissues increased 1.7 to 3.5 times in comparison with that in normal lung tissues. The increase in tissue content of glycosaminoglycans was accompanied by an increase in the chondroitin sulfate level in every histologic type of lung cancer, as well as marked increase in hyaluronic acid level in squamous cell carcinomas, and a moderate increase in small cell carcinomas. The concentrations of dermatan sulfate and heparan sulfate in lung cancer tissues did not show any significant changes compared with those in normal lung tissues. The increase in total amount and changes in the composition of glycosaminoglycans in human lung cancer tissue were closely related to the histologic type of the tumor. In adenocarcinomas, some acid glycoprotein with sialic acid was simultaneously detected during the separating course of glycosaminoglycans, which was considered to be derived from mucinous substances related to adenocarcinoma cells.

Bioaccumulation of ergovaline in bovine lateral saphenous veins in vitro1,2

Ergot alkaloids have been associated with vasoconstriction in grazing livestock affected by the fescue toxicosis syndrome. Previous in vitro investigations studying how ergot alkaloids caused vasoconstriction have shown that ergovaline has a distinct receptor affinity and sustained contractile response. A similar contractile response has not been noted for lysergic acid. The objectives of this study were to determine if repetitive in vitro exposure of bovine lateral saphenous vein to lysergic acid or ergovaline would result in an increasing contractile response and if a measurable bioaccumulation of the alkaloids in the vascular tissue occurs over time. Segments of vein were surgically biopsied from healthy, Angus x Brangus cross-bred, fescue-naïve yearling heifers (n = 16) or collected from healthy mixed breed and sex cattle immediately after slaughter (n = 12) at a local abattoir. Veins were trimmed of excess fat and connective tissue, sliced into cross-sections, and suspended in a myograph chamber containing 5 mL of oxygenated Krebs-Henseleit buffer (95% O(2)/5% CO(2); pH = 7.4; 37 degrees C). Contractile responses to repetitive additions of ergovaline (1 x 10(-9) and 1 x 10(-7) M) and lysergic acid (1 x 10(-5) and 1 x 10(-4) M) were evaluated using the biopsied veins. For the bioaccumulation experiments, veins collected at the abattoir underwent repetitive additions of 1 x 10(-7) M ergovaline and 1 x 10(-5) M lysergic acid and the segments were removed after every 2 additions and media rinses for alkaloid quantification via HPLC/mass spectrometry. Contractile data were normalized as a percentage of contractile response induced by a reference dose of norepinephrine (1 x 10(-4) M). Repetitive additions of 1 x 10(-9) M ergovaline and 1 x 10(-5) and 1 x 10(-4) M lysergic acid resulted in contractile response with a negative slope (P < 0.02). In contrast, repetitive addition of 1 x 10(-7) M ergovaline resulted in a contractile response that increased with each addition (P < 0.01). Lysergic acid and ergovaline were detected at all 4 exposure levels (2x to 8x), but only the 1 x 10(-7) M ergovaline treatment resulted in increased tissue content as the number of exposures increased (P < 0.05). These data indicate that ergovaline, but not lysergic acid, bioaccumulates with repetitive exposure in vitro. These results suggest that ergovaline may have a greater potential for inducing toxicosis in grazing animals than lysergic acid because of its potential to bioaccumulate at the cellular site of action.