Viscum album prevents haematological changes,electrolyte imbalance,changes in liver function enzymes and histological alterations in some selected tissues in cadmium chloride-intoxicated rats

BackgroundThe use of Viscum album to treat different diseases is popular in the practise of alternative medicine. We investigated the ability of the aqueous extract of V. album to protect against the toxic effects of cadmium.MethodsThirty rats used for the experiment were treated as follows; Group 1 – no cadmium or extract. Group 2–10 mg/kg body weight of cadmium chloride. Group 3–10 mg/kg body weight of cadmium chloride and 200 mg/kg body weight of aqueous extract of V. album. Group 4–10 mg/kg body weight of cadmium chloride and 400 mg/kg body weight of aqueous extract of V. album. Group 5–10 mg/kg body weight of cadmium chloride with 800 mg/kg body weight of aqueous extract of V. album. Group 6–10 mg/kg body weight of cadmium chloride and atorvastatin (100 mg/kg body weight).ResultsApart from WBC and platelets, other haematological parameters and electrolytes, urea and creatinine levels were not significantly affected by the administration of cadmium chloride along with the aqueous extract of V. album. Treatment with the extract caused significant decreases in the hepatosomatic index, cardiosomatic index, and increase in renosomatic index of the test rats. It also resulted in significant (P < 0.05) decrease in AST level. Histological report also shows that treatment with the extract restored the normal myocardium and vascular architecture of the heart, normal portal and vascular architecture of the liver and normal glomerular and tubular architecture of the kidney, in the cadmium-intoxicated experimental rats.ConclusionV. album protects against the toxic effects of cadmium chloride.     

Influence of the Main Phospholipid (MPL) from Thermoplasma Acidophilum and of Liposomes from MPL on Living Cells: Cytotoxicity and Mutagenicity

Abstract

Liposomes of the main phospholipid (MPL) from the archaebacterium Thermplasma acidophilum were investigated for their interference with living cells. Growth of mouse lymphoma cells L5178Y, permanent hamster fibroblasts V79, Ehrlich-mouse-ascites tumor (EMAT) cells and a variety of other celltypes was not influenced by these liposomes. Mutagenicity and antimutagenic efficacy were tested with Salmonella typhimurium TA100 in the “Ames plate-incorporation test”. No cytotoxicity and mutagenicity of liposomes from MPL was detected. The influence of MPL liposomes on ion transport, intracellular pH, electrolytes, membrane potential, energy metabolism, and the biosynthesis of proteins and nucleic acids in EMAT cells is demonstrated in detail.     

Toxicity and Immunogenic Properties of Liposomal form of Vipera Libetina Venom

Abstract

The incorporation of Vipera libetina venom into liposomes obtained from pure egg phosphatidyl choline by the reverse phase evaporation method decreases its toxicity by 3-fold - in mice LD₅₀ for the native toxin is 2.22 mg/kg body weight and for the liposomal toxin 6.9 mg/kg. Subcutaneous injection of liposomal preparation into mice stimulates the development of cellular immunity and reproduces the reaction of the delayed-type hypersensitivity. It is. also, shown that after a single dose immunization of mice with liposomes containing 1xLD₅₀ dose of the venom, the.titer.of antibodies increases at the early postinfection period and is maintained on high level longer than after the injection of the native venom. Thus, liposom.es can be succesfully used for antiserum production and protective immunization against Vipera libetina venom.     

Effects of copper and other metals on fertilization,embryo development,larval survival and settlement of the polychaete Nereis (Neanthes) virens

Summary

Nectochaete larvae of the ecologically and economically important ragworm, Nereis virens, were exposed to cadmium, chromium, copper, lead and zinc dissolved in seawater to nominal concentrations ranging from 0 to 5000 μg l^?1. Copper was the most toxic (mean LC50 of 76.5 μg l^?1 ± 95% CI 73.8–79.2 after 96 h exposure) and so was used for subsequent experiments. Exposure of gametes to greater than 500 μg l^?1 copper for 2 or 4 h at 10°C prior to fertilization, or a 10 min exposure during fertilization, significantly reduced embryo developmental success. The effect of copper on larval settlement was also assessed using sediment spiked to a range of concentrations (0, 50, 250, 500, 1000 mg kg!1 dry weight). Significantly fewer larvae were found in sediment of $250 mg kg!1 in comparison to the control or the 50 mg kg!1 treatment. Assessment of living larvae also confirmed a significant reduction in settlement, but in all treatments compared to the control, although the number of dead larvae also increased as the concentrations increased. These effects may have important implications for reproductive success and recruitment of N. virens to polluted sediments.     

Subcutaneous administration of bovine superoxide dismutase protects lungs from radiation-induced lung injury

Abstract

Background. The objective of the present study was to determine whether single administration of the antioxidant enzyme bovine superoxide dismutase (bSOD) after radiation therapy (RT) mitigates development of pulmonary toxicity in rats. Methods. Female F344 rats (n = 60) were divided among six experimental groups: (1) RT, single dose of 21 Gy to the right hemithorax; (2) RT + 5 mg/kg bSOD; (3) RT + 15 mg/kg bSOD; (4) No RT; (5) sham RT + 5 mg/kg bSOD; and (6) sham RT + 15 mg/kg bSOD. A single subcutaneous injection of bSOD (5 or 15 mg/kg) was administered 24 h post-radiation. The effects of bSOD on radiation-induced lung injury were assessed by measurement of body weight, breathing frequency, and histopathological changes. Immunohistochemistry was used to evaluate oxidative stress (8-OHdG⁺, NOX4⁺, nitrotyrosine⁺, and 4HNE⁺ cells), macrophage activation (ED1⁺), and expression of profibrotic transforming growth factor-β or TGF-β in irradiated tissue. Results. Radiation led to an increase in all the evaluated parameters. Treatment with 15 mg/kg bSOD significantly decreased levels of all the evaluated parameters including tissue damage and breathing frequency starting 6 weeks post-radiation. Animals treated with 5 mg/kg bSOD trended toward a suppression of radiation-induced lung damage but did not reach statistical significance. Conclusions. The single application of bSOD (15 mg/kg) ameliorates radiation-induced lung injury through suppression of reactive oxygen species/reactive nitrogen species or ROS/RNS-dependent tissue damage.     

Hypoglycemic activity of the fungi Cordyceps militaris, Cordyceps sinensis, Tricholoma mongolicum, and Omphalia lapidescens in streptozotocin-induced diabetic rats

Crude extracts were prepared from fruiting bodies and mycelia of the medicinal fungus Cordyceps militaris, and a polysaccharide-enriched fraction was obtained after extraction with hot water and ethanol precipitation. Polysaccharide-enriched fractions were similarly prepared from Cordyceps sinensis, Omphalia lapidescens, and Tricholoma mongolicum. The various aforementioned preparations were orally administered into different groups of adult rats 24 h before an intraperitoneal injection of streptozotocin (40 mg/kg body weight), and subsequently daily for another 4 days. The dosage used was 10 mg/kg body weight for polysaccharide-enriched preparations and 100 mg/kg body weight for crude extracts. Control rats received distilled water instead of crude extract or polysaccharide-enriched preparation. It was found in the control rats that plasma glucose level rose from about 90 mg/dl before streptozotocin injection to levels that were maintained at about 300 mg/dl postinjection. All preparations produced hypoglycemic effects. C. militaris polysaccharide-enriched fraction displayed a more prominent effect than that of C. sinensis polysaccharide-enriched fraction which in turn was more potent than that of O. lapidescens and T. mongolicum polysaccharide-enriched fractions. The hypoglycemic effect of C. militaris polysaccharide-enriched fraction was dose-dependent.     

Further studies of the effect of L-canavanine on the tobacco hornworm, Manduca sexta.

Fifth instar Manduca sexta growth response to injected doses of canavanine was concentration-dependent over a range of 0·5 to 2·0 mg/g body weight. Twenty-four hr after injection of ¹⁴C-guanidinooxy-d,l-canavanine, M. sexta larvae incorporated approximately 3·6% of the labelled l-canavanine into protein of non-gut tissue. Adult M. sexta mortality was related to the level of injected canavanine over a range of 2 to 8 mg/g body weight. Injection of as little as 2 mg canavanine/g body weight caused hyperactivity in adult M. sexta. Arginine, able to negate the toxic effects of canavanine during larval growth, was only marginally capable of overcoming canavanine effects on larval-pupal ecdysis.     

Effects of alpha-chlorohydrin on the male reproductive tissues of the Polynesian rat,Rattus exulans

Abstract

Doses of α-chlorohydrin (‘Epibloc’) were administered by gavage to mature male Polynesian rats (Rattus exulans) at 100, 200, and 300 mg per kg body weight. Animals that survived were sacrificed either 1 day or 7 days later for assessment of epididymal and testicular cytology and sperm viability. Two of 10 animals died 6 days after treatment with 100 mg/kg; 1/6 died within 24 h of treatment with 200 mg/kg, though 6/10 died when left for 7 days; 300 mg/kg was lethal to all 3 rats tested. After 1 day, microscopic lesions were observed in the Initial Segment of the epididymis of 4/6 rats dosed with 100 mg/kg and in all 5 of the 200 mg/kg group; however, in only one animal at the higher dose level was the damage severe enough to cause epithelial exfoliation and potential blockage of the lumen. In all the animals that survived for 7 days testicular and epididymal cytology were normal, and viable spermatozoa were present at all levels of the tract. Autopsies revealed no evidence of gross epididymal lesions in any of the animals that died from the drug. We conclude that although α-chlorohydrin causes minor lesions in the epididymis of this feral species, the damage appears to be reversible in animals that survive an acute dose, and the drug cannot be considered an effective chemosterilant, as distinct from a poison.     

Non‐specific immunity and disease resistance are enhanced by the polysaccharide fraction of a marine chlorophycean macroalga in Oreochromis niloticus (Linnaeus, 1758)

Polysaccharides (PF) from marine macroalgae, Caulerpa scalpelliformis were extracted and tested for its potential immunostimulatory and disease resistance properties in fish. Five groups of Nile tilapia (n = 6), Oreochromis niloticus (Linnaeus, 1758) were intraperitoneally administered with the different doses of PF (2, 20 or 200 mg/kg body weight) or with yeast‐derived commercial immunostimulant, Macrogard^? (20 mg/kg body weight), to compare the effectiveness. An untreated control group was also maintained. A total of fifteen fibre reinforced plastic tanks (150 L, ambient temperature and light conditions) were used, with triplicate tanks for each group. Only four fish per tank (totally 12 fish from a group) were taken at random and assayed. PF enhanced all the tested non‐specific serum immune responses namely lysozyme, myeloperoxidase, antiprotease, and bactericidal activities. There was an upregulation of the genes encoding IL‐1β, lysozyme and TNF‐α in the spleen of PF injected fish as compared to the control group. In order to study the overall functional immunity, disease resistance test was conducted. Another five groups of fish (n = 10) were treated by intraperitoneal injection with different doses of PF or Macrogard^? or untreated as mentioned earlier in triplicates (30 fish per group in three tanks, totally 150 fish in 15 tanks). Seven days post treatment, fish were challenged by intraperitoneal administration of live virulent Aeromonas hydrophila. PF treated fish were protected with significant reduction in the mortality and the consequent increased relative percent survival (RPS) of 92 in the least (2 mg/kg) and middle dose (20 mg/kg) groups. The disease resistance experiment was repeated again but this time, fish were challenged 21 days post treatment that resulted in RPS of 50 for the middle dose. The results clearly show that the intraperitoneal administration of the polysaccharide fraction had a stimulating effect on the non‐specific immune responses, immune gene expression and disease resistance.     

Toxic cyanobacteria in Greek freshwaters

Cyanobacterial scums, collected in 1987 from four Greek freshwater lakes, were examined for their toxicity to mice. Species ofMicrocystis, Oscillatoria, Anabaenopsis, andAnabaena were dominant in the samples. All samples tested had toxic effects on mice after intraperitoneal injection. The lethal dose (LD₅₀) ranged from 40 to 1500 mg cyanobacterial dry weight kg^?1 body weight and gross pathological signs of poisoning were characteristic of cyanobacterial hepatotoxins. The toxicities of the Greek cyanobacterial blooms were similar to those reported for blooms elsewhere in the world, shown to be responsible for the poisoning of wild and domestic animals.     

Localization of retinal dehydrogenase type 1 in the stomach and intestine

Rats were injected with liposomes containing iodixanol (CTP10 Injection; 100 mg iodine per kg body weight) followed by a second injection of 125I-tyramine-cellobiose-albumin microspheres. The amounts of phagocytosed and degraded labelled albumin in liver were measured. A reduced uptake and degradation of albumin microspheres was observed when the labelled microspheres were injected 2 h or 24 h after the liposomes compared with that obtained in control animals receiving saline. No effect on the uptake and degradation of labelled microspheres was observed when the time lag between the injection of liposomes and labelled microspheres was 1 week. The data show that the uptake and degradation of 125I-tyramine-cellobiose-albumin microspheres can be used as indicators of Kupffer cell phagocytotic function following drug uptake by these cells.     

Pyrene degradation and copper and zinc uptake by Fusarium solani and Hypocrea lixii isolated from petrol station soil

Aims: This study aimed to isolate and identify potential polycyclic aromatic hydrocarbon (PAH)‐degrading and/or metal‐tolerant fungi from PAH‐contaminated and metal‐contaminated soils. Methods and Results: Pyrene‐degrading fungi were isolated from contaminated soil and tested for metal (Cu, Zn and Pb) compound solubilization and metal accumulation. Three strains of Fusarium solani and one of Hypocrea lixii were able to degrade more than 60% of initial supplied pyrene (100 mg l^?1) after 2 weeks. The isolates were grown on toxic metal (Cu, Pb and Zn)‐containing media: all isolates accumulated Cu in their mycelia to values ranging from c. 5·9 to 10·4 mmol per kg dry weight biomass. The isolates were also able to accumulate Zn (c. 3·7–7·2 mmol per kg dry weight biomass) from zinc phosphate‐amended media. None of the isolates accumulated Pb. Conclusions: These fungal isolates appear to show promise for use in bioremediation of pyrene or related xenobiotics and removal of copper and zinc from wastes contaminated singly or in combination with these substances. Significance and Impact of the Study: Microbial responses to mixed organic and inorganic pollution are seldom considered: this research highlights the abilities of certain fungal strains to interact with both xenobiotics and toxic metals and is relevant to other studies on natural attenuation and bioremediation of polluted sites.     

Effect of hypoglycemia on the brain free fatty acid level and the uptake of fatty acids by phospholipids

The effect of hypoglycemia on the uptake of [1-¹⁴C]arachidonate and [1-¹⁴C]oleate into a synaptosomal and microsomal glycerophospholipids was investigated. In the presence of ATP, Mg²⁺ and CoA, rat brain synaptosomes and micorsomes catalyze the transfer of arachidonate and oleatc into glycerophospholipids. Arachidonate was mainly incorporated into phosphatidylinositol (PI) and phosphatidylcholine (PC), whereas oleate was incorporated into phosphatidylcholine and phosphatidylethanolamine (PE).Hypoglycemia was produced by intraperitoneal injection of 10 or 100 units of crystalline insulin per kg body weight. Two hours after injection the blood glucose level decreased to 10–20 mg%. The content of brain phospholipids was slightly decreased but the change was not statistically significant. The level of free fatty acids (FFA) was increased. More pronounced and reproducible changes were found when hypoglycemia was produced by injection of 100 units of insulin per/kg body weight. Changes in brain cortex were similar to those observed in microsomes and synaptosomes. Hypoglycemia affected the incorporation of arachidonic acid into glycerophospholipids of brain membranes. Uptake of [1-¹⁴C]arachidonate was decreased selectively by 50% (into phosphatidic acid /PA/) when hypogiycemia was produced by injection of 10 units of insulin per kg body weight. The Higher dose of insulin 100 units per kg body weight produced a 20% inhibition of arachidonate incorporation into synaptosomal PI and a 13% decrease of incorporation into microsomal phosphatidylcholine. Incorporation of [1-¹⁴C]oleate into membrane phospholipids was not changed by hypoglycemic insult. It is proposed that the disturbances in fatty acid level, particularly arachidonate, and decreased uptake of arachidonic acid by synaptosomal glycerophospholipids may be responsible for alteration of membrane function and changes of synaptic processes.     

Antivitamin B-6 effect of 1-aminoproline on rats

By intraperitoneal injection of 1-aminoproline, death after severe convulsion was observed in rats (LD₅₀ of 1-amino-l-proline, 26 mg per kg of body weight for young male rats fed a normal diet). The vitamin B-6-deficient rats were more sensitive to this hydrazino acid than the normal rats. The toxic effect was completely prevented by the administration of pyridoxine. 1-Amino-d-proline was less toxic than the l-isomer. By the 1-aminoproline treatment, the most remarkable changes in the free amino acid levels were the striking increases in the concentrations of α-aminodipic acid, citrulline and cystathionine in all the tissues tested, except in brain. Some unidentified ninhydrin-positive substances appeared. These results indicate that 1-aminoproline greatly disturbed the amino acid pattern, i.e. the amino acid metabolism in rats.     

Grazing on toxic cyanobacterial blooms by tadpoles of edible frog Rana grylio

Tadpoles of Rana grylio were raised as edible frogs in fishponds of Guanqiao in Wuhan City, Hubei, China, during cyanobacterial blooms from June to October. The dominant cyanobacterial species was Microcystis, which was found to be lethally toxic by intraperitoneal (i.p.) mouse bioassay. Little is known about the effect of tadpoles on toxic cyanobacterial blooms. To evaluate the potential of the tadpoles to graze on cyanobacterial blooms, the tadpoles were fed on Microcystis collected from the field in the laboratory. The Microcystis cells decreased from 1.19 × 10⁷ cells mL^?1 to 3.23 × 10⁶ cells mL^?1, with a sharp reduction of 73% of the initial Microcystis population observed in the first 24 h after introduction of the tadpoles. The ponds containing tadpoles had a markedly lower density of Microcystis than those lacking tadpoles. Tadpoles exposed to either cultured Microcystis aeruginosa (NIES–90, 2.768 µg microcystins mg^–1 dw^–1) cells or lysed M. aeruginosa cells grew well, however, indicating that they were unaffected by Microcystis toxins. We found a significant increase in tadpole body weight after feeding on either field Microcystis or cultured M. aeruginosa. The mean increase in individual body weight was 20 mg day^?1 when fed on Microcystis from the pond, and 7 mg day^?1 when fed on M. aeruginosa from culture. Our study strongly suggested that there is a direct trophic relationship between R. grylio tadpoles and toxic Microcystis blooms and they possess the potential to graze on toxic Microcystis. The results imply that R. grylio tadpoles may play an important ecological role in reducing toxic cyanobacterial blooms caused by Microcystis.     

Estimation of Excess Cancer Risk on Time-Weighted Lifetime Average Daily Intake of PAHs from Food Ingestion

The purposes of this study were to quantify the time-weighted, lifetime average, daily intake (LADI) of polycyclic aromatic hydrocarbons (PAHs) through food ingestion and to estimate the excess cancer risk based on lifetime dietary PAH intake. Twenty-seven different food commodities were selected from the 2001 Korean National Health and Nutrition survey based on their frequent consumption and high PAH level. The foods were analyzed for the profile of 14 PAH congeners using high performance liquid chromatography (HPLC) and fluorescence detector. Considering the toxic equivalent (TEQ) level converted with the toxic equivalent factors (TEFs), the highest total TEQ level of PAHs in foods was detected from roasted laver at 1.2 ug TEQ/kg. For the PAH exposure assessment according to ingested foods, the average body weight was separated according to the following age groups, 1–6, 7–19, 20–64 and over 64 years, and the daily food ingestion rates from the National Health and Nutrition survey were used. The estimated Lifetime Average Daily Intake (LADI) of PAHs was 3.22 × 10^–3 ug/kg/day for carcinogenic effects and was higher in the younger age groups under 20 years old than in the older groups. The dietary excess cancer risk estimated using the cancer potency of benzo(a)pyrene (7.3(mg/kg/day)^?1) was 2.3 × 10^?5, which is equivalent to a probability of tumor eruption in the upper gastrointestinal tract of two per hundred thousand persons.     

Protective effect of Zizyphus lotus jujube fruits against cypermethrin-induced oxidative stress and neurotoxicity in mice

Context: Cypermethrin (CYP) is a synthetic pyrethroid insecticide used worldwide in agriculture, home pest control. The toxicity of CYP is well studied in many organisms.

Objective: The aim of present study was to investigate the protective effect of Zizyphus lotus (Zizyp) fruit against neurotoxicity and oxidative stress induced by CYP in mice.

Materials and methods: Mice were divided into four groups of six each: groups I and II were used as control and CYP control (20?mg/kg body weight). While, groups III was orally treated with Zizyphus lotus fruit (5?g/kg body weight) plus CYP (20?mg/kg body weight) for 18?days. Furthermore, HPLC–ESI–MS–MS (Q-Tof) and GC–MS were used to identify the compounds fraction.

Results: Antioxidant enzyme catalase (CAT), neurotoxicity enzyme acetylcholinesterase (AChE) activities and hydrogen peroxide (H₂O₂)_, malondialdehyde (MDA) levels were determined in the liver, kidney and heart. CYP caused decreased CAT activity, inhibition of AChE activity and increased the levels of H₂O₂ and MDA in heart, liver and kidney.

Conclusion: Our results indicate that Zizyp fruit is markedly effective in protecting mice against CYP-induced biochemical changes. This protection may be due to its antioxidant property and scavenging ability against active free radicals.     

人脐带间充质干细胞重复静脉注射动物毒性试验

摘要 目的：评价多次尾静脉注射脐带间充质干细胞（hUC-MSCs）对小鼠的体内毒性作用。方法：48只健康ICR小鼠,按性别和体重随机分为4组（即对照组、低剂量组、中剂量组和高剂量组）。小鼠通过微静脉注射不同剂量hUC-MSCs悬浮液,间隔3天给药1次,共给药4次。记录小鼠摄食量、体重、体温,给药结束后恢复两周后牺牲动物作大体解剖,检查各个器官器质性病变;利用流式细胞仪分别检测CD3、CD4、CD8阳性细胞亚群数量;ELISA试剂盒检测血清IgM、IgG、C3、C4指标;对肺脏、脾脏、肾脏行组织病理学检查。结果：实验组与对照组相比较,注射不同剂量干细胞后一般观察、体重、体温、摄食量、IgM以及C3在给药期和恢复期均未发生显著变化。在恢复期,注射中、高剂量hUC-MSCs组血清IgG和C4水平略有降低,但未达到显著水平P<0.05;CD4阳性T细胞集群数量以及CD4/CD8系数在hUC-MSCs中、高剂量组显著上升（P<0.05）。大体剖检,除脾脏相比溶媒对照组略显增大外其它各器官均未发现肉眼可见明显异常;称重后发现hUC-MSCs高剂量组脾重量与溶媒对照组相比显著升高（P<0.05）。脾脏、肺脏、肾脏病理学检测未见明显异常。结论：健康ICR小鼠尾静脉注射临床剂量hUC-MSCs（1×10⁶ cells/kg）可能调动动物免疫反应,此外,未观察到hUC-MSCs对小鼠有明显毒副作用。     

Investigation of liver binding of pentachlorophenol based upon measurement of protein adducts

Abstract

Covalent binding of reactive metabolites of pentachloropheno (PCP) was investigated both in vitro andin vivo in the livers of male Sprague-Dawley rats via measurement of protein adducts. Cysteinyl adducts of quinones andsemiquinones in liver cytosolic (Cp) andnuclear (Np) proteins were assayed after catalytic cleavage by Raney nickel. Results from in vitro experiments confirmed that PCP metabolism produced tetrachlorobenzoquinones andthe comsponding tetrachlorobentosemiquinones which subsequently bound to sulphydryl groups in liver proteins. In vivo, the production of cysteinyl adducts increased with the administered dosage (0–40 mg PCP per kg body weight) andpresented evidence of saturable metabolism. Results suggest two metabolic pathways for PCP, including a high-affinity low-capacity pathway anda low-affinity high-capacity pathway. Time-course experiments in vivo andin vitro suggested that quinone adducts partlcipated in multiple substitution reactions with protein and/or non-protein thiols, andpointed to possible formation of protein-protein cross-links in vivo. The elimination rate constants of quinone adducts in vitro were about 0.35 h^?1 in liver Cp. The elimination of quinone adducts in vivo appeared to follow biphasic kinetics with rate constants for the terminal phase being 0.014 and0.008 h^?1 in liver Cp andNp, respectively.