Plant Protection with Inorganic Ions

Gradients of salts of the specific ion repellents for Meloidogyne incognita -- NH₄⁺, K⁺, Cl⁻, and NO₃⁻ -- have been demonstrated to shield tomato roots from infestation in soil. The strategy of these greenhouse experiments was to interpose a salt barrier in a soil column between the plant roots and the nematodes. The relative effectiveness of the salts as a barrier to infective second-stage juveniles in a sandy loam was NH₄NO₃, NH₄Cl > KNO₃ > KCl. Some of these ions are beneficial to plant growth, and the results suggest that a new environmentally tolerable means of plant protection is possible.     

Biochemical Oxidation of Reduced-form Cytochrome c by Mercuric Ions

A simple method has been developed for DNA isolation from purified chloroplasts of Marchantia polymorpha L. (liverwort) cell suspension cultures. Purified chloroplasts exhibited ribulose-bisphosphate carboxylase activity comparable to that of Fraction 1 protein obtained from Nicotiana tabacum. Fraction 1 protein isolated from purified chloroplasts clearly showed large and small subunits when subjected to isoelectric focussing. These results indicate that the purified chloroplasts are intact. DNA isolated from purified chloroplasts showed a covalently closed circular form, and restriction endonuclease digestions of the chloroplast DNA showed clear fragmentation indicating that the DNA was sufficiently free from those of other organelles.     

Dietary Supplementation of Selenium in Inorganic and Organic Forms Differentially and Commonly Alters Blood and Liver Selenium Concentrations and Liver Gene Expression Profiles of Growing Beef Heifers

In geographic regions where selenium (Se) soil concentrations are naturally low, the addition of Se to animal feed is necessary. Even though it is known that Se in grass and forage crops is primarily present in organic forms (especially as L-selenomethionine, L-selenocystine, and L-selenocystathionine), the feeding of Se in the naturally occurring organic selenium (OSe) compounds produces higher blood and tissue Se levels than the inorganic Se (ISe) salts, and that animal metabolism of OSe and ISe is fundamentally different. Se is commonly added in inorganic form as sodium selenite to cattle feeds because it is a less expensive source of supplemental Se then are OSe forms. A trial was conducted with growing cattle to determine if the addition of OSe versus ISe forms of Se in beef cattle feed produces differences in hepatic gene expression, thereby gaining insight into the metabolic consequence of feeding OSe versus ISe. Thirty maturing Angus heifers (261?±?6 days) were fed a corn silage-based diet with no Se supplementation for 75 days. Heifers (body weight?=?393?±?9 kg) then were randomly assigned (n?=?10) and fed Se supplements that contained none (control) or 3 mg Se/day in ISe (sodium selenite) or OSe (Sel-Plex®) form and enough of a common cracked corn/cottonseed hull-based diet (0.48 mg Se/day) to support 0.5 kg/day growth for 105 or 106 days. More Se was found in jugular whole blood and red blood cells and biopsied liver tissue of ISe and OSe treatment animals than control animals, and OSe animals contained more Se in these tissues than did ISe. Microarray and bioinformatic analyses of liver tissue gene expression revealed that the content of at least 80 mRNA were affected by ISe or OSe treatments, including mRNA associated with nutrient metabolism; cellular growth, proliferation, and immune response; cell communication or signaling; and tissue/organ development and function. Overall, three Se supplement-dependent gene groups were identified: ISe-dependent, OSe-dependent, and Se form-independent. More specifically, both forms of supplementation appeared to upregulate mitochondrial gene expression capacity, whereas gene expression of a protein involved in antiviral capacity was downregulated in ISe-supplemented animals, and OSe-supplemented animals had reduced levels of mRNA encoding proteins known to be upregulated during oxidative stress and cancerous states.     

Mercury Toxicity and the Mitigating Role of Selenium

Mercury is a well-known environmental toxicant, particularly in its most common organic form, methylmercury. Consumption of fish and shellfish that contain methylmercury is a dominant source of mercury exposure in humans and piscivorous wildlife. Considerable efforts have focused on assessment of mercury and its attendant risks in the environment and food sources, including the studies reported in this issue. However, studies of mercury intoxication have frequently failed to consider the protective effects of the essential trace element, selenium. Mercury binds to selenium with extraordinarily high affinity, and high maternal exposures inhibit selenium-dependent enzyme activities in fetal brains. However, increased maternal dietary selenium intakes preserve these enzyme activities, thereby preventing the pathological effects that would otherwise arise in their absence. Recent evidence indicates that assessments of mercury exposure and tissue levels need to consider selenium intakes and tissue distributions in order to provide meaningful risk evaluations.     

Pregnancy Alters Renal and Blood Burden of Mercury in Females

The concentrations of copper and zinc in the tissues of alcohol-addicted people can significantly correlate with the variables describing their mental state. Studies on the homeostasis of zinc in alcohol-dependent patients have often been characterized by low hypozincemia detection. This may be caused by a low content of zinc in blood serum (1%) compared to the average zinc level in the body. Unfortunately, most authors have identified extracellular zinc in their studies. In the available literature, data on the level of copper in patients suffering from alcohol dependence are inconsistent. Our study included 100 alcohol-addicted patients (the study group) and 50 healthy subjects (the control group). Mental state was measured using appropriate psychometric scales. We used inductively coupled plasma mass spectrometry (ICP-MS) to determine copper and zinc content. Our results confirm the purposefulness of the use of zinc concentration in erythrocytes as a diagnostic parameter for low zinc status in alcohol-dependent patients. Alcohol-dependent patients with reduced concentrations of zinc in erythrocytes/copper in blood plasma differed significantly from alcohol-dependent patients with normal concentrations in terms of clinical parameters. With regard to zinc in blood plasma and copper in erythrocytes, this situation has not been found. The clinical symptoms of hypozincemia and copper deficiency in patients addicted to alcohol usually relate to disorders in central nervous system functioning, and they result in a decreased quality of physical and mental life.     

Selenium Supplementation Alters Gene Expression Profiles Associated with Innate Immunity in Whole-Blood Neutrophils of Sheep

Footrot (FR) is a common, contagious bacterial disease of sheep that results in lameness and significant economic losses for producers. We previously reported that sheep affected with FR have lower whole-blood (WB) selenium (Se) concentrations and that Se supplementation in conjunction with routine control practices accelerates recovery from FR. To determine whether oral Se-yeast administered at supranutritional levels (>4.9 mg Se/week) alters the ability of sheep to resist or recover from FR infection, 60 ewes with and 60 ewes without FR were drenched once weekly for 62.5 weeks with 0, 4.9, 14.7, or 24.5 mg organic Se-yeast (30 ewes per treatment group). Footrot prevalence and severity were measured at 0, 20, 28, 40, and 60 weeks of Se supplementation. Genomic expression of eight WB-neutrophil genes for selenoproteins and seven WB-neutrophil genes for proteins involved in innate immunity was determined at the end of the treatment period using SYBR Green and quantitative polymerase chain reaction methodology. Supranutritional Se-yeast supplementation successfully increased Se status in sheep but did not prevent FR. Supranutritional Se-yeast supplementation increased WB-neutrophil expression of genes involved in innate immunity: l-selectin, interleukin-8 receptor, and toll-like receptor 4, which were or tended to be lower in ewes affected with FR. Furthermore, supranutritional Se-yeast supplementation altered the expression of selenoprotein genes involved in innate immunity, increasing selenoprotein S and glutathione peroxidase 4 and decreasing iodothyronine deiodinases 2 and 3. In conclusion, supranutritional Se-yeast supplementation does not prevent FR, but does alter WB-neutrophil gene expression profiles associated with innate immunity, including reversing those impacted by FR.     

Repetitive Microstimulation Alters the Cortical Representation of Movements in Adult Rats

In order to examine the effects of repetitive stimulation on functional cortical organization, standard intracortical microstimulation (ICMS) techniques were used to generate maps of movement representations in motor cortex of rat. After identification of caudal and rostral forelimb fields and adjacent vibrissae and neck fields, one or more representational borders were defined in greater detail. Then a microelectrode was introduced into one of these representational fields, and ICMS current pulses were delivered at a rate of 1/sec for 1 to 3 hr. Following repetitive ICMS, significant changes in movement representations were observed using current levels that were either suprathreshold or subthreshold for evoking the site-specific movement. Electromyographic activity could be evoked at suprathreshold and near-threshold current levels, but not at the subthreshold current levels used here. Significant border shifts ranged from 210 to 670 μm. In each case in which shifts occurred, there appeared to be expansion of the movement represented at the repetitively stimulated site. The effects were progressive and reversible. These results suggest that at least under these unusual experimental circumstances, large representational changes can be generated very rapidly within motor cortex in the absence of any evident peripheral feedback.     

The Uptake of Inorganic Ions by Plant Leaves

A technique for estimating the gross uptake of ions by plantleaves is described. Uptake of strontium-89 and caesium-137from droplets applied to plant leaves was greatly affected bythe humidity of the air, the effect being markedly dependenton the initial concentration of the solution. Uptake was greaterin light than in darkness; the effect of temperature was relativelysmall. The percentage uptake of the nuclides was reduced only to arelatively small extent as the concentration of carrier ionwas increased. This was in marked contrast to the situationwhen leaves were fully immersed in solution. ¹³⁷Cs was takenup to a greater extent than iodine, phosphate, or sulphate.The role of exchange and of fixed negative charges in uptakeis discussed.     

啤酒酵母菌对无机硒的有机转化

利用啤酒酵母菌对无机硒(亚硒酸钠)进行有机转化。通过在培养基中加入不同浓度的无机硒溶液和不同时间加入无机硒溶液,于28℃、220 r/min摇床条件下培养5 d,离心得菌细胞,测定前样品预处理:破碎菌细胞,显微镜下计数,计算破碎率,破碎后的菌体装入透析袋于蒸馏水中透析除去无机硒。准确测定无机硒,用浓硫酸-高氯酸的消化体系消化样品后,紫外分光光度法于335 nm处测量吸光度,在标准曲线上查出硒含量,计算无机硒的转化率。啤酒酵母菌的最佳加硒时间为24 h,亚硒酸钠浓度大于12μg/mL对啤酒酵母菌转化无机硒有明显抑制作用,啤酒酵母菌对无机硒的摄入率约为62%,转化率约为53%;超生波细胞粉碎仪破碎细胞的破碎率为55%左右。结果表明,啤酒酵母菌可以转化无机硒。     

Valproic Acid Alters GnRH-GABA Interactions in Cycling Female Rats

Summary of the aims Women with epilepsy using antiepileptic drug valproic acid (VPA) often suffer from reproductive endocrine disorders, menstrual disorders and polycystic ovaries. Valproic acid exerts anticonvulsive effects via gamma amino butyric acid (GABA) neurotransmitter system, which also acts as a neurochemical regulator of gonadotropin-releasing hormone (GnRH) neurons and suggests possibility of valproic acid mediated interruption in gonadotropin releasing hormone pulse generator in hypothalamus. The aim of this study was to investigate the effects of valproic acid treatment on the expression of gonadotropin releasing hormone, gamma amino butyric acid and polysialylated form of neural cell adhesion molecule (PSA-NCAM) a marker of neuronal plasticity in the median preoptic area (mPOA) and median eminence-arcuate (ME-ARC) region having GnRH neuron cell bodies and axon terminals, respectively. Methods Three-month-old virgin Wistar strain female rats received VPA (i.p.) at a dose of 300 mg/kg once a day for 12 weeks; control group received an equivalent volume of vehicle. GnRH, GABA and PSA-NCAM expressions were studied by immunohistofluorescence technique from mPOA and ME-ARC region of hypothalamus. Ovarian histology was also studied using Mayer’s Haematoxylin-Eosin staining method. Results GnRH and PSA-NCAM staining was much higher in mPOA and ME-ARC region from vehicle treated control proestrous rats, whereas VPA treatment significantly enhanced GABA expression, and reduced both GnRH and PSA-NCAM expression. Mayer’s Haematoxylin-Eosin staining of mid-ovarian sections revealed significantly higher number of ovarian follicular cysts in VPA treated rats. Conclusions Our findings of alterations in GnRH and GABA expression and GnRH neuronal plasticity marker PSA-NCAM as well as changes in ovarian histology suggest that treatment with VPA disrupts hypothalamo-hypophyseal-gonadal axis (HPG) at the level of GnRH pulse generator in hypothalamus.     

蒙药孟根乌苏-18 味丸、孟根乌苏炮制品的肝肾毒性作用研究

目的:对比研究孟根乌苏(水银)-18味丸、孟根乌苏(水银)炮制品与硫化汞、氯化汞、氯化亚汞的肝肾毒性作用。方法:将Wistar雄性大鼠,根据体重随机分为正常对照组、孟根乌苏-18味丸低、高剂量组(0.29,2.9 g·kg~(-1)·d~(-1))、孟根乌苏-18味丸简化方组(0.26 g·kg~(-1)·d~(-1))、孟根乌苏炮制品低、高剂量组(0.033,0.33 g·kg~(-1)·d~(-1))、硫化汞组(17.39 mg·kg~(-1)·d~(-1))、氯化汞组(4.06 mg·kg~(-1)·d~(-1))、氯化亚汞组(35.3 mg·kg~(-1)·d~(-1)),共9组,每组6只。各组大鼠适应7天后,灌胃给药7天后分别检测肝肾功能,肝肾组织形态学变化,并用电感耦合等离子体发射光谱仪(ICP-OES)和电感耦合等离子体质谱仪(ICP-MS)法测肾汞蓄积量,原位末端标记(TUNEL)法测肾细胞凋亡,免疫组化法测肾Ⅲ型胶原蛋白表达,实时荧光定量PCR(Real-Time-PCR)法检测肾脏MT-1、MT-2基因表达的变化。结果:在实验过程中,氯化亚汞组4只大鼠死亡,因此未对实验数据进行统计。大鼠连续7天给药后,药物对各组大鼠肝、肾功能并无影响。肝脏和肾脏病理检查结果表明,孟根乌苏-18味丸和孟根乌苏炮制品低剂量组肝细胞肿胀变性程度较轻,肾小球轻度肥大,肾小管上皮轻度肿胀变性,孟根乌苏-18味丸和孟根乌苏炮制品高剂量组以及硫化汞组大鼠肝脏、肾脏均出现了一定的病理变化,而氯化汞、氯化亚汞组大鼠肝肾病理变化更显著。与正常对照组和孟根乌苏炮制品低剂量组相比,氯化汞组大鼠肾汞蓄积量显著升高(P0.01);硫化汞、氯化汞组大鼠肾细胞凋亡率和Ⅲ型胶原蛋白表达显著升高(P0.01);与正常对照组比较,氯化汞组大鼠肾组织中MT-1和MT-2 m RNA表达显著升高(P0.01,P0.05);氯化汞组大鼠肾组织中MT-1表达显著高于孟根乌苏炮制品低剂量组(P0.01)。结论:临床常用量的孟根乌苏-18味丸和孟根乌苏炮制品肝肾毒性要远低于氯化亚汞、氯化汞。     

Selenium and Mercury in Pelagic Fish in the Central North Pacific Near Hawaii

Protective effects of selenium against mercury toxicity have been demonstrated in all animal models evaluated. As interactions between selenium and mercury and their molar ratios in seafood are essential factors in evaluating risks associated with dietary mercury exposure, considering mercury content alone is inadequate. In this study, the absolute and molar concentrations of mercury and selenium were determined in edible portions from 420 individual fish representing 15 species of pelagic fish collected from the central North Pacific Ocean near Hawaii. Selenium was in molar excess of mercury in almost all fish species evaluated. The rank order of mean Se/Hg molar ratios was striped marlin (17.6) > yellowfin tuna (14.1) > mahimahi (13.1) > skipjack tuna (12.8) > spearfish (11.4) > wahoo (10.8) > sickle pomfret (6.7) > albacore tuna (5.3) > bigeye tuna (5.2) > blue marlin (4.1) > escolar (2.4) > opah (2.3) > thresher shark (1.5) > swordfish (1.2) > mako shark (0.5). With a Se/Hg molar ratio of less than 1, mako shark was the only fish containing a net molar excess of mercury. A selenium health benefit value based on the absolute amounts and relative proportions of selenium and mercury in seafood is proposed as a more comprehensive seafood safety criterion.     

Effects of Lycopene and Proanthocyanidins on Hepatotoxicity Induced by Mercuric Chloride in Rats

To evaluate the protective potential of lycopene (Lyc) and proanthocyanidins (PCs) against mercuric chloride (HgCl₂)-induced hepatotoxicity, the study focused on the mechanism of oxidative stress. Firstly, the rats were subcutaneously (s.c.) injected with 0, 2.2, 4.4, and 8.8 μmol/kg HgCl₂. Additionally, 40 mg/kg Lyc and 450 mg/kg PCs were given to the rats intragastrically (i.g.) before exposure to 8.8 μmol/kg HgCl₂. Then, body weight, liver weight coefficient, mercury (Hg) contents, histological feature, ultrastructure, apoptosis, reactive oxygen species (ROS), glutathione (GSH), glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and malondialdehyde (MDA) in the liver were measured. Lactate dehydrogenase (LDH) and alanine transaminase (ALT) in serum were determined. After exposure to different concentrations of HgCl₂,it was found that Hg contents, pathological and ultrastructure injury, activities of LDH and ALT, apoptosis, and levels of ROS, GSH, and MDA increased and the activities of SOD and GSH-Px decreased in a concentration-dependent manner. Further investigation found that pretreatment with Lyc and PCs inhibited ROS production, protected antioxidant enzymes, and reversed hepatotoxicity. We concluded that Lyc and PCs had hepatoprotective effects on HgCl₂-induced toxicity by antagonizing oxidative stress in rat liver.     

Community Analysis of a Mercury Hot Spring Supports Occurrence of Domain-Specific Forms of Mercuric Reductase

Mercury is a redox-active heavy metal that reacts with active thiols and depletes cellular antioxidants. Active resistance to the mercuric ion is a widely distributed trait among bacteria and results from the action of mercuric reductase (MerA). Protein phylogenetic analysis of MerA in bacteria indicated the occurrence of a second distinctive form of MerA among the archaea, which lacked an N-terminal metal recruitment domain and a C-terminal active tyrosine. To assess the distribution of the forms of MerA in an interacting community comprising members of both prokaryotic domains, studies were conducted at a naturally occurring mercury-rich geothermal environment. Geochemical analyses of Coso Hot Springs indicated that mercury ore (cinnabar) was present at concentrations of parts per thousand. Under high-temperature and acid conditions, cinnabar may be oxidized to the toxic form Hg²⁺, necessitating mercury resistance in resident prokaryotes. Culture-independent analysis combined with culture-based methods indicated the presence of thermophilic crenarchaeal and gram-positive bacterial taxa. Fluorescence in situ hybridization analysis provided quantitative data for community composition. DNA sequence analysis of archaeal and bacterial merA sequences derived from cultured pool isolates and from community DNA supported the hypothesis that both forms of MerA were present. Competition experiments were performed to assess the role of archaeal merA in biological fitness. An essential role for this protein was evident during growth in a mercury-contaminated environment. Despite environmental selection for mercury resistance and the proximity of community members, MerA retains the two distinct prokaryotic forms and avoids genetic homogenization.     

Supplemented Organic and Inorganic Selenium Affects Milk Performance and Selenium Concentration in Milk and Tissues in the Guanzhong Dairy Goat

Trace amounts of selenium (Se) are essential for several organisms, and deficiencies therein have adverse effects on growth, development, and reproduction; this is particularly significant in animals raised for milk and livestock production. To study the effect of Se on Guanzhong dairy goats, their diets were supplemented with different sources (inorganic or organic) and Se concentrations (0.2 or 0.4 mg Se/kg). A non-Se-fortified basal diet served as a negative control, and a sixth treatment group received both inorganic and organic Se sources (0.2 mg Se/kg diet each). Dietary Se supplementation increased milk production, with organic Se being more effective than inorganic Se. Selenium supplementation also increased Se concentration and glutathione peroxidase activity in whole blood, with organic Se more effective than inorganic Se at the same Se concentration. With increasing Se in diets, the Se content in milk increased markedly, reaching a plateau value at day 30 in all groups, and organic Se (0.4 mg/kg diet) had the best effect. In addition, dietary Se sources and concentrations markedly affected Se concentrations in different tissues and organs. Thus, organic Se supplementation of a basal diet at 0.4 mg/kg is practically applicable for Se-enriched milk and meat production in Guanzhong dairy goats.