Chemical analysis of tobacco cigarette for organochlorine insecticides and heavy metal composition

The effects of cigarette smoking on smokers and the environment were investigated by analyzing some cigarette brands available to Nigerian smokers for the presence of some of the widely used organochlorine (OC) pesticides and heavy metals. OC pesticide residues were extracted from tobacco smoke, separated and identified using column chromatographic and thin layer chromatographic techniques respectively, while ashed and unashed commercial cigarettes were analyzed for heavy metals – Cr, Mn, Ni, Pb, Zn, Cd, Cu, and Co using atomic absorption spectrometry. TLC data indicated that dieldrin and p,p′-DDD were present in some of the cigarette brands. The results of heavy metal analysis showed that the levels of some toxic heavy metals and pollution index were higher in unashed cigarettes than corresponding ashes. Evidence suggests that significant amounts of these toxic metals are inhaled by the smoker and/or released into the environment in the process of cigarette smoking, thus confirming that cigarette smoking is harmful to the environment and human health.     

Plant extract reduces tobacco smoke harmful effects on alveolar macrophage immune responses

Tobacco smoke is a major factor responsible for lung cancer and chronic obstructive pulmonary disease. Although the best solution to reduce the incidence of these diseases is to quit smoking, there are still a large number of smokers. Thus, given the immunoregulatory properties of plant extracts, their capacity to reduce tobacco smoke harmful effects on alveolar macrophage (AM) functions was investigated. AM were treated with tobacco smoke extract and parenchymata tissue extract (PTE), or mesophyll cell extract (MCE) of Spinacia oleacea. The effects of tobacco smoke extract from PTE and MCE-treated cigarette filters were also investigated. AM production of tumor necrosis factor (TNF), interleukin-10 (IL-10) and macrophage chemoattractant protein-1 (MCP-1), and AM cytotoxicity were measured. Tobacco smoke extract significantly inhibited TNF, IL-10, and MCP-1 release, and AM cytotoxicity. The addition of PTE and MCE to tobacco smoke extract abrogated the inhibition of AM mediator release. However, only MCE restored AM cytotoxicity. Interestingly, tobacco smoke extract of PTE and MCE-treated cigarette filters showed reduced effects on AM functions. Tobacco smoke extract from MCE-treated (0.25%) cigarette filters did not inhibit TNF, IL-10, and MCP-1 release in contrast to tobacco smoke extract from buffer-treated cigarette filters. AM cytotoxic activity was not inhibited by the treatment with tobacco smoke extract from MCE-treated cigarette filters. Our data suggest that the presence of plant extract in cigarette filters reduces the inhibitory effects of cigarette smoke on AM functions. Thus, MCE-treated cigarette filters may help reducing lung diseases associated with smoking.     

Correlation of cadmium and aluminum in blood samples of kidney disorder patients with drinking water and tobacco smoking: related health risk

The combined exposure to aluminum (Al) and cadmium (Cd) causes more pronounced adverse health effects on humans. The kidneys are the main organs affected by internal exposure to Cd and Al via food and non-food items. The objective of present study was to measure the Al and Cd concentrations in cigarettes tobacco (branded and non-branded) and drinking water (domestic treated, ground and lake water) samples in southern part of Pakistan, to assess the risk due to ingestion of water and inhalation of cigarettes smoke containing high concentrations of both elements. The study population (kidney disorder and healthy) divided into two group based on consuming lake and ground water, while smoking non-branded cigarette as exposed, while drinking domestic treated water and smoking branded cigarette as non-exposed. Electrothermal atomic absorption spectrometry was used to determined Cd and Al concentrations in tobacco, drinking water and blood samples. The resulted data indicated that the levels of Al and Cd in lake and underground water were higher than the permissible limit in drinking water recommended by the World Health Organization. The biochemical parameters of exposed and referent patients, especially urinary N-acetyl-h-glucosaminidase, were used as a biomarkers of kidney disorder. Exposed kidney disorder patients have higher levels of Cd and Al than the exposed referents subjects, while difference was significant when compared to resulted data of non-exposed patients and referents (p = 0.01–0.001). The pearson correlation showed positive correlation between both toxic element concentrations in water, cigarettes versus blood samples of exposed subjects (r = 0.20–0.67 and 0.71–0.82), while lower values were observed for non-exposed subjects (r = 0.123–0.423 and 0.331–0.425), respectively.

     

Degradation of Benzo[a]Pyrene in Soil with Arbuscular Mycorrhizal Alfalfa

Mycorrhizal and non-mycorrhizal alfalfa (Medicago sativa) was grown in pots containing soil artificially contaminated with various levels of benzo[a]pyrene (B[a]P)(0, 1, 10 and 100 mg kg^–1). Soil and plants were sampled after 30, 40, 50, 60 and 90 days and compared with unlanted pots. The percentage of mycorrhizal root length colonized by Glomus caledoniun was not significantly affected by the addition of B[a]P up to 10 mg kg^–1 but was significantly lower at 100 mg kg^–1B[a]P compared with low concentrations (p < 0.05). There was no difference in soil polyphenol oxidase and dehydrogenase activity among the controls and applications of 1 and 10 mg kg^–1 of B[a]P. However, enzyme activities were significantly higher at 100 mg kg^–1B[a]P compared with the other three treatments, and there was no mycorrhizal effect. Over a period of 90 days the concentration of B[a]P in soil in which alfalfa was grown was significantly lower than in unplanted soil (p < 0.05). Degradation rates of B[a]P added at 1, 10 and 100 mg kg^–1 without G. caledonium were 76, 78 and 53%, and with mycorrhizal inoculation were 86, 87 and 57%. The degradation rate in unplanted soil was significantly lower than in planted soil, and was significantly higher in medium- and low-B[a]P treatments than in the high B[a]P concentration tested. There is a possibility of enhancement phytoremediation of PAHs in rhizosphere soil with arbuscular mycorrhizal fungi.     

Multiple-time contamination of benzo(a)pyrene in soils inhibits soil enzymatic activities

Benzo[a]pyrene (B[a]P) is accumulating in soils in a low-dose cumulative manner. The objectives of this study were to investigate the effects of B[a]P on the extractable and available fractions of B[a]P and on soil enzymatic activity using multiple-time superimposed and one-time contamination approaches. Results showed that the contents of B[a]P rapidly decreased in the first 14?d and later decreased slowly from 14 to 56?d in both one-time and multiple-time contamination tests. The contents of B[a]P in the multiple-time contamination test were lower than those in the one-time test. Soil urease, sucrase and dehydrogenase activities were rapidly inhibited in the early stage (14?d) and stimulated during the rest of the incubation, and soil dehydrogenase activity was more sensitive to B[a]P contamination than the other enzymes. High concentrations of B[a]P in soil led to greater inhibition of enzymatic activity than that at low concentrations in the early period of culture. Soil enzyme activities were weakly inhibited in multiple-time compared with in one-time contamination tests and were lower in the subsurface layer than in the surface layer. Our results revealed that the multiple-time superimposed approach might be better than one-time contamination for evaluating B[a]P risk in soil.     

Evaluation of 3-hydroxy-benzo[a]pyrene levels in Nile tilapia (Oreochromis niloticus) after waterborne exposure to Benzo[a]pyrene

3-Hydroxy-benzo[a]pyrene (3-OH-BaP), a toxic compound with the ability to covalently bind with the macromolecules (proteins and DNA), is one of the major phenolic metabolites of benzo[a]pyrene (BaP). The purpose of this study was to evaluate the presence of 3-OH-BaP in the bile and plasma of Nile tilapia by HPLC with fluorescence detection, after waterborne exposure to BaP (10 and 100?µg?L^?1). Metabolites were detected in bile and plasma, and conjugates of 3-OH-BaP (glucuronide and/or sulphate conjugates) were the majority forms in both biological fluids, glucuronide 3-OH-BaP being the main metabolite in bile. Our data suggest that extrahepatic tissues as intestine or gill are important in BaP metabolism and need to be the considered sources of metabolites released into the blood. Although, low levels of 3-OH-BaP in toxic form (free form) were detected in plasma, one cannot exclude the possibility of circulating the levels leading to adverse effects.     

Mutagenicity of polyaromatic hydrocarbons by chemical models for cytochrome P450 in Ames assay

DNA damage is an important step in carcinogenesis. The Ames assay is a short-term screening of carcinogens that induce DNA damage. Most carcinogens require enzymatic activation through oxidation by cytochrome P450 (CYP450) in the presence of S9 mix. A combination of iron (Fe)(III) porphyrin and an oxidant is also able to oxidize compounds as an alternative metabolic pathway to CYP450. Previously it was reported that a chemical model containing a water-soluble 5,10,15,20-tetrakis(1-methylpyridinium4-yl)porphyrinatoiron(III) chloride (4-MPy) and tert-butyl hydroperoxide (t-BuOOH) activated aromatic amines and amides. In this study, a chemical model composed of an Fe porphyrin, water-insoluble 5,10,15,20-tetrakis(pentafluorophenyl)porphyrinatoiron(III) chloride (F₅P) or water-soluble 4-MPy was optimized with an oxidant – t-BuOOH, magnesium monoperoxyphthalate (MPPT), or iodosylbenzene (PhIO). Subsequently the mutagenicity of benzo[a]pyrene (B[a]P) and chrysene in Salmonella typhimurium TA strains was compared. B[a]P was activated by a combination of F₅P or 4-MPy plus MPPT or PhIO in S. typhimurium TA1538. The B[a]P-induced mutagenicity with F₅P plus oxidant was higher than 4-MPy plus oxidant. Mutagenicity of chrysene, a tetracyclic aromatic hydrocarbon, was not detected in the presence of F₅P/PhIO in S. typhimurium TA98, but was activated in the presence of F₅P/MPPT. The F₅P/MPPT activated other polyaromatic hydrocarbons (PAH) in the S. typhimurium TA98 assay including dibenz[a,c]anthracene, dibenz[a,h]anthracene, 3-methylcholanthrene, and benzo[a]anthracene. The results indicated that the F₅P/MPPT was the most efficient model for detecting PAH-induced mutagenicity in the Ames assay.     

Impaired phospholipid metabolism in Saccharomyces cerevisiae by increased phospholipase B activity induced by 4-(methyl nitrosamino)-1-(3-pyridyl)-1-butanone

Tobacco products and cigarette smoke cause many respiratory diseases including cancer. 4-(Methyl nitrosamino)-1-(3-pyridyl)-1-butanone (NNK) is a potent carcinogen in cigarette smoke, but its effect on lipid metabolism remains enigmatic. Hence, Saccharomyces cerevisiae was exposed to different concentrations of NNK (0–400?µmol?L^?1) to elucidate its role in lipid metabolism. Exposure to NNK substantially decreases (about 60%) of the phospholipid content with a concomitant increase in lysophospholipids. Significant reduction was observed in the phosphatidylcholine followed by phosphatidylethanolamine with NNK-treated cells. On the contrary, cells accumulated significant amount of neutral lipids and free fatty acids. Exposure of yeast cells (wild-type cells and three plbΔ mutant strains) to NNK greatly enhances the hydrolysis of phospholipid in the presence of calcium. We are the first to report that exposure to NNK enhances phospholipase B (PLB), particularly plb1p activity. Furthermore, NNK also promotes the alteration of phospholipid fatty acid (FA) content. These results suggest that NNK aids in the degradation of phospholipids by enhanced PLB activity and is accompanied with FA alteration. Understanding the altered phospholipid metabolism in the presence of NNK remains a worthy pursuit.     

栘依树抗氧化剂对香烟烟气自由基及人淋巴细胞姐妹染色单体交换率的影响

采用电子顺磁共振法研究了栘依树抗氧化剂对香烟烟气自由基含量的影响,并比较了不同自由基含量的香烟烟气冷凝物(CSC)对人淋巴细胞姐妹染色单体的损伤情况.在香烟滤嘴中注入50μL0.2%的栘依树抗氧化剂溶液后,香烟烟气中气相、粒相自由基含量分别降低40.2%和28.3%,平均降低39.6%;中高剂量的实验烟(添加栘依树),与对照烟(不加栘依树)相比,人淋巴细胞姐妹染色单体交换率显著降低(t＞t0．05),其中中剂量烟降低幅度更大;低剂量实验烟与对照烟相比,姐妹染色单体交换率也有降低趋势,但差异不显著(t相似文献   

Influence of marine alga against mercury-induced biochemical changes,structural destabilization,and genotoxicity in marine tiger fish,Therapon jarbua

This study elucidated the protective role of alga against mercury (Hg)-induced toxicity in marine fish. Bath immersion with mercury chloride (HgCl) [0.125 and 0.25?ppm] in combination with the algal extract (3?ppm) to Therapon jarbua offered a significant protection against Hg only induced disturbed liver function, damaged histoarchitecture, elevated oxidative stress, and DNA fragmentation of tissues. Mercury exposure decreased hepatic superoxide dismutase (SOD), catalase (CAT) activities, and the level of nonprotein-soluble thiol (NPSH, GSH), with a concomitant increase in thiobarbituric acid reactive substances (TBARS) in the gill, kidneys, liver, and blood. Algal supplementation diminished the rise in TBARS restoring CAT, SOD, and GSH levels to control. Reduced generation of free radicals may be correlated to protect DNA stability and morphology. This study indicates the ability of alga to ameliorate Hg-mediated injuries. In conclusion, Kappaphycus alvarezii extract did not show any toxicity and its safety is suggestive for using as a supplement in fish food.     

Aspects of iron and nitrogen nutrition in the red tide dinoflagellateGymnodinium sanguineum

Iron-stress-mediated effects on biochemical constituents of the red tide dinoflagellateGymnodinium sanguineum Hirasaka were examined in 1988 by comparing Fe-replete and Fe-deplete batch cultures. The influence of nitrogen source (NO₃ or NH₄) on characteristics of Fe-deplete cells was also studied [i.e., Fe-deplete/NO₃-grown (= — Fe/NO₃) vs Fe-deplete/NH₄-grown (= — Fe/NH₄)]. Common to both N sources were reductions of chlorophylla (chla) and Fe quotas (per cell volume) by 75% and ca. 1.5 orders of magnitude, respectively, under Fe depletion. The Fe requirement ofG. sanguineum exceeded those of certain neritic diatoms by one to two orders of magnitude. — Fe/NH₄ cells exhibited 30 to 50% greater N quotas and free amino acid:protein ratios than did Fe-deplete cells grown on NO₃. In vivo fluorescence:chla increased with Fe deficiency particularly in — Fe/NO₃ cultures, surpassing — Fe/NH₄ values by ca. two-fold. Effects of Fe depletion were consistent with this element's essential role in the biosynthesis of chla and components of the photosynthetic electron transport (PET) system, and also in NO₃ utilization. Fe:N ratios were larger (1.5-fold) for iron-deficient NO₃-grown than NH₄-grown cells, likely reflecting the Fe content of NO₃ assimilatory enzymes [nitrate (NR) and nitrite (NiR) reductase] and of electron transport components needed to provide reductant, coupled with a diminished capacity of — Fe/NO₃ cells to acquire and assimilate nitrogen. Indicators of PET efficiency suggested that under iron stress, supply of Fe for NR and NiR is partly at the expense of iron-containing PET components. Utilization of nitrate by NO₃-grown cells was inhibited sufficiently by Fe depletion to yield symptoms bordering on N deficiency. In an ecological context, the most important effect mediated by nitrogen source may be the determination of critical Q_Fe (i.e., Fe required to just sustain maximal growth), thereby regulating the degree of growth limitation for a given subsaturating iron concentration.     

2种典型家用消毒剂对水生生物的急性毒性影响及生态效应阈值研究

家用消毒剂大量用于日常生活中,进入水体环境后对水生生物产生潜在危害效应,目前尚缺乏保护水生生物安全的生态效应阈值。本研究以2种典型家用消毒剂(有效成分分别为对氯间二甲苯酚和次氯酸钠,前者命名为消毒剂A、后者为消毒剂B)为研究对象,开展其对8种不同营养级淡水水生生物的急性毒性效应研究。结果表明,除底栖动物外,消毒剂B对藻类、溞类和鱼类的急性毒性均高于消毒剂A;我国本土种稀有鮈鲫对2种消毒剂的敏感性高于其他2种鱼类;2种消毒剂对藻类的毒性高低均为近头状伪蹄形藻斜生栅藻蛋白核小球藻;近头状伪蹄形藻对2种消毒剂最敏感。基于上述毒性数据构建了物种敏感分布(SSD)曲线,计算对保护95%的物种不受影响时所对应的污染物浓度(HC5),并结合评估因子法推导出2种消毒剂预测无效应浓度(PNEC)值作为急性生态效应阈值,消毒剂A和消毒剂B的PNEC值分别为13.16 mg·L~(-1)(有效成分对氯间二甲苯酚PNEC值为0.33 mg·L~(-1))和0.71 mg·L~(-1)(有效成分次氯酸钠PNEC值为0.01 mg·L~(-1)),消毒剂A对淡水生物的PNEC比消毒剂B大了一个数量级,表明相较于消毒剂B,消毒剂A对水生态环境更为友好。本研究结果可为制订典型家用消毒剂的水质基准提供科学依据。     

Bioaccumulation and adverse effects of trace metals and polycyclic aromatic hydrocarbons in the common onion Allium cepa as a model in ecotoxicological bioassays

The common onion Allium cepa can be easily used in ecotoxicological bioassays to evaluate the bioavailability and adverse effects of pollutants and complex mixtures like waste, industrial effluent or coal-mining drainage that contain elevated levels of trace metals and polycyclic aromatic hydrocarbons (PAHs). In this study, onions were exposed for 7 days to individual metals (1?µg?mL^?1), i.e., aluminium, copper, iron and manganese, or PAHs (1.5?ng?mL^?1), i.e., benzo[a]antracene and benzo[a]pirene. Biological effects, measured as growth inhibition of roots and leaves, were integrated with analysis of bioaccumulation in roots, bulbs and leaves. Copper, iron and benzo[a]pirene caused a significant inhibition in root development of newly formed tissues, whereas only slight variations were caused by other chemicals; the number of new root filaments and the length of the leaves did not show significant variations, thus not representing sensitive parameters to evaluate adverse effects of pollutants in A. cepa. Chemicals bioaccumulation was always significant in roots, whereas levels in bulbs and leaves exhibited increased levels only for manganese, and a decrease for aluminium. The overall results confirmed the sensitivity of the bioassays with A. cepa, suggesting their utility for future applications to evaluate the adverse effects of complex mixtures containing metals or PAHs.     

吸烟烟气引起鼠肝微粒体脂质过氧化的ESR波谱研究

本文用ESR自旋捕集技术,以4-POBN为捕集剂,研究了吸烟烟气处理的鼠肝微粒体体系中的自由基性质。结果发现,在吸烟烟气处理的鼠肝微粒体中,4-POBN捕集到了自由基信号,这一信号的强度随温育时间的延长及充入烟气的量增加而增强,根据其性质及ESR波谱参数分析,体系中产生了脂类自由基。结果说明,吸烟烟气刺激了鼠肝微粒体的脂质过氧化。     

Concentrations of selected metals in blood,serum, and urine among US adult exclusive users of cigarettes,cigars, and electronic cigarettes

Data from National Health and Nutrition Examination Survey for 2013–2014 were used to compare observed levels of selected metals in blood, serum, and urine among US adults aged ≥20 years for exclusive cigar, cigarettes, and e-cigarette users. Adjusted geometric means for e-cigarette users were found to be higher than for cigar users for blood manganese (10.3 vs. 7.9 µg/L, p = 0.02). Cigar users were found to have lower adjusted geometric means than cigarette users for urine cobalt (0.22 vs. 0.4 µg/L, p = 0.04) and urine antimony (0.03 vs. 0.06 µg/L, p = 0.03). Adjusted levels of blood selenium, serum copper, selenium, and zinc, and urine arsenic, barium, molybdenum, tin, strontium, thallium, tungsten, and uranium were found to be comparable among cigarettes only, cigar only, and electronic cigarettes only users. However, irrespective of the comparative levels of these metals among cigar, cigarette, and e-cigarette users, focus must be to assess the short- and long-term health effects of the exposure to these metals particularly nanoparticles via inhalation from e-cigarette aerosols.     

Daily rhythm of O2-evolution in the cyanobacterium Trichodesmium thiebautii under natural and constant conditions

The rate of oxygen evolution by the tropical marine cyanobacterium Trichodesmium thiebautii was recorded at different times during the day in samples collected in 1992 from the Bahama Islands and the NE Caribbean Sea. This cyanobacterium is unique in that it is the only non-heterocystous diazotroph capable of N₂-fixation in daylight. Oxygen evolution was measured under conditions of natural day/night (LD, N=50), constant light (LL, N=14), and constant dark (DD, N=2×14). Photosynthesis vs intensity (P-I) relationships were calculated at various times of day, and the following parameters were used for further evaluation: photosynthesic capacity (P _max, 66 to 91 mg O₂ mg chl a ^-1 h^-1), initial slope of the P-I curve (, 0.23 to 0.27), dark respiration (R, 12 to 27 mg O₂ mg chl a ^-1 h^-1), and the intensity at which O₂ consumption is compensated by O₂ production (I _c, 78 to 160 Em^-2 s^-1). All means showed large standard deviations (for some parameters more than 200%). In some cases, these variations could be explained with a sinusoidal 24-h time course, but only the compensation point showed a significant daily variation (p0.001) in both LD and DD. The fact that the time course of I _c typical for natural conditions remains rhythmic under constant dark conditions strongly suggests a circadian regulation. Few circadian rhythms have been observed in prokaryotes, and this appears to be the first observation of such a rhythm in a cyanobacterium which fixes N₂ in daytime.     

Benzoic acid, a stimulant of odorant receptors of Bombyx mori, is rapidly metabolized to N-benzoylserine on the antennae

Summary. Benzoic acid is a known stimulant of olfactory receptor cells in trichoid sensilla on the antennae of adult females of Bombyx mori. Exposure of freshly excised antennae, from B. mori females and males, to vapours of [¹⁴C]-benzoic acid revealed that the adsorbed acid is rapidly metabolized (15% conversion in 10 s), suggesting the presence of a highly active enzymatic system. The major product could be identified as N-benzoylserine by use of [²H₅]-benzoic acid, and mass spectrometry linked to a gas chromatograph (GC-MS) or a liquid chromatograph (LC-MS), for analysis of the metabolites. The conjugation of benzoic acid with an amino acid apparently represents a novel mode for the rapid deactivation of odorant molecules. Received 26 March 2001.     

Protection of Genetic Diversity and Maintenance of Connectivity among Reef Corals within Marine Protected Areas

Abstract: High‐latitude coral reefs (HLRs) are potentially vulnerable marine ecosystems facing well‐documented threats to tropical reefs and exposure to suboptimal temperatures and insolation. In addition, because of their geographic isolation, HLRs may have poor or erratic larval connections to tropical reefs and a reduced genetic diversity and capacity to respond to environmental change. On Australia's east coast, a system of marine protected areas (MPAs) has been established with the aim of conserving HLRs in part by providing sources of colonizing larvae. To examine the effectiveness of existing MPAs as networks for dispersal, we compared genetic diversity within and among the HLRs in MPAs and between these HLRs and tropical reefs on the southern Great Barrier Reef (GBR). The 2 coral species best represented on Australian HLRs (the brooding Pocillopora damicornis and the broadcast‐spawning Goniastrea australensis) exhibited sharply contrasting patterns of diversity and connectedness. For P. damicornis, the 8‐locus genetic and genotypic diversity declined dramatically with increasing latitude (N_a= 3.6–1.2, H_e= 0.3–0.03, N_g:N = 0.87–0.06), although population structure was consistent with recruitment derived largely from sexual reproduction (G_o:G_e= 1.28–0.55). Genetic differentiation was high among the HLRs (F_ST[SD]= 0.32 [0.08], p < 0.05) and between the GBR and the HLRs (F_ST= 0.24 [0.06], p < 0.05), which indicates these temperate populations are effectively closed. In contrast for G. australensis, 9‐locus genetic diversity was more consistent across reefs (N_a= 4.2–3.9, H_e= 0.3–0.26, N_g:N = 1–0.61), and there was no differentiation among regions (F_ST= 0.00 [0.004], p > 0.05), which implies the HLRs and the southern GBR are strongly interconnected. Our results demonstrate that although the current MPAs appear to capture most of the genetic diversity present within the HLR systems for these 2 species, their sharply contrasting patterns of connectivity indicate some taxa, such as P. damicornis, will be more vulnerable than others, and this disparity will provide challenges for future management.     

Neuroprotective potential of N -acetylcysteine in carbofuran neurotoxicity: A biochemical and histopathological study

The present study was undertaken to evaluate biochemical markers of chronic carbofuran exposure to rats in terms of lipid peroxide and intrasynaptosomal calcium levels and to correlate them with the histopathological changes in brain regions. A significant increase in lipid peroxidation (LPO) in terms of thiobarbituric acid reactive substances (TBARS) was observed in the cerebral cortex (65%) and brain stem (33%) after carbofuran exposure. This was accompanied by a significant increase (87%) in the intracellular free-Ca²⁺ [Ca²⁺]_i levels. N-acetylcysteine (NAC) administration, on the other hand, reversed the carbofuran-induced increase in LPO and [Ca²⁺]_i. Histopathological studies of carbofuran-exposed brain revealed high frequency of pyknotic neurons in the cerebral cortex and microhaemorrhages in the brain stem. NAC supplementation to carbofuran-treated animals resulted in normalisation of the brain architecture as seen by a reduction in the number of pyknotic nuclei in the cerebral cortex. These findings indicate that increased LPO and elevated [Ca²⁺]_i levels are involved in the development of carbofuran neurotoxicity and are eventually responsible for the pathological alterations. The study also demonstrates potential neuroprotective effect of NAC treatment in carbofuran neurotoxicity.     

Cloning of urea transporters from the kidneys of two batoid elasmobranchs: evidence for a common elasmobranch urea transporter isoform

One of the two phloretin-sensitive, facilitated urea transporters identified from the kidneys of the myliobatiform, euryhaline elasmobranch, Dasyatis sabina, a 379 amino acid protein ([D. sabina]strUT-2), was very similar to the 380 amino acid isoform (shUT) present in the kidney of the squaliform, dogfish shark, Squalus acanthias (a species that can be considered marginally euryhaline since it utilizes upper estuarine, as well as ocean habitats). To test the proposal that this isoform is a conserved urea transporter (UT) expressed in the kidneys of diverse elasmobranchs, UTs were cloned from the kidneys of a rajiform elasmobranch, the stenohaline skate, Leucoraja ocellata and another dasyatid stingray, the marginally euryhaline, Dasyatis say. Utilizing 5′/3′ RACE, a 2,060 nt cDNA that encoded a phloretin-sensitive, 378 amino acid skate urea transporter ([L. ocellata]skUT-2) and a 1,683 nt cDNA that encoded a stingray 379 amino acid UT ([D. say]strUT-2) were obtained. These deduced UTs have a very high sequence identity with the known elasmobranch Uts. [L. ocellata]skUT-2 was 86% identical to [D. sabina]strUT-2 and 84% identical to [S. acanthias]shUT. [D. say]strUT-2 was 97% identical to the [D. sabina]strUT-2. These findings support the hypothesis that a conserved UT isoform is present in the kidneys of marine elasmobranchs, and is an important pathway for facilitated urea transport in the kidneys of marine elasmobranchs. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. H.A. Gefroh and E.E. Cwengros contributed equally to this study.