Cotyledonary responses to maternal selenium and dietary restriction may influence alterations in fetal weight and fetal liver glycogen in sheep

To examine the effects of maternal supranutritional selenium (Se) and nutrient restriction during mid and late gestation on placental characteristics and fetal liver glycogen, ewes received either adequate Se (ASe) or high Se (HSe) prior to breeding. On d 64 of gestation, ASe and HSe ewes remained at 100% of requirements (controls; CON) or were restricted (RES; 60% of requirements). On d 135 of gestation, fetal weight (P ≤ 0.08) was greatest in both HSe and CON ewes. Placentome number, mass, and caruncular and cotyledonary weight were not different (P ≥ 0.17) among treatments. Fetal mass:placental mass ratio was less (P = 0.06) in RES compared to CON ewes. Compared to ASe, HSe exhibited increased (P ≤ 0.08) cellular proliferation and DNA concentration and decreased (P = 0.07) cellular size in cotyledonary tissue. Nutritional restriction decreased (P ≤ 0.08) cotyledonary protein concentration and cellular size. VEGF receptor 1 (Flt) mRNA in cotyledonary tissue was greater in HSe compared with ASe ewes (P = 0.06) and in RES compared with CON ewes (P = 0.08). There was no effect of diet on caruncular growth variables (P ≥ 0.13) or on placental vascularity (P ≥ 0.11). Progesterone was greater (P ≤ 0.08) in ASe–RES ewes compared to all groups at d 90 and ASe–CON and HSe–CON at d 104. Although fetal glucose and cortisol concentrations were not affected by diet, fetal liver glycogen was greater (P = 0.04) in ASe–RES compared to ASe–CON and HSe–RES ewes with HSe–CON being intermediate. Both Se and nutritional plane may impact placental function and fetal growth, as fetal weight and liver glycogen are altered despite similar placental vascularity measurements.     

Plasma levels of trace elements and exercise induced stress hormones in well-trained athletes

This study analyzed the variation and relationship of several trace elements, metabolic substrates and stress hormones activated by exercise during incremental exercise. Seventeen well-trained endurance athletes performed a cycle ergometer test: after a warm-up of 10 min at 2.0 W kg⁻¹, the workload was increased by 0.5 W kg⁻¹ every 10 min until exhaustion. Prior diet, activity patterns, and levels of exercise training were controlled, and tests timed to minimize variations due to the circadian rhythm. Oxygen uptake, blood lactate concentration, plasma ions (Zn, Se, Mn and Co), serum glucose, non-esterified fatty acids (NEFAs) and several hormones were measured at rest, at the end of each stage and 3, 5 and 7 min post-exercise. Urine specific gravity was measured before and after the test, and participants drank water ad libitum.Significant differences were found in plasma Zn and Se levels as a function of exercise intensity. Zn was significantly correlated with epinephrine, norepinephrine and cortisol (r = 0.884, P < 0.01; r = 0.871, P < 0.01; and r = 0.808, P = 0.05); and Se showed significant positive correlations whit epinephrine and cortisol (r = 0.743, P < 0.05; and r = 0.776, P < 0.05). Neither Zn nor Se levels were associated with insulin or glucagon, and neither Mn nor Co levels were associated with any of the hormones or substrate metabolites studied. Further, while Zn levels were found to be associated only with lactate, plasma Se was significantly correlated with lactate and glucose (respectively for Zn: r = 0.891, P < 0.01; and for Se: r = 0.743, P < 0.05; r = 0.831, P < 0.05).In conclusion, our data suggest that there is a positive correlation between the increases in plasma Zn or Se and stress hormones variations induced by exercise along different submaximal intensities in well-hydrated well-trained endurance athletes.     

Trace mineral status and liver and blood parameters in sheep without mineral supply compared to local roe deer (Capreolus capreolus) populations

General health, clinical-chemical blood analysis and copper (Cu), zinc (Zn), selenium (Se) and vitamin E concentrations in plasma and liver tissue (wet weight, ww) of two extensive grazing sheep flocks without mineral supply were compared to the status of local roe deer (Capreolus capreolus) populations (liver samples). Both sheep flocks were classified as healthy except for a remarkable variation in body weight and a slight foot rot infection in one flock. Hematology of sheep was normal, and total protein and creatinine as well as activities of creatin kinase, aspartat-amino-transferase, alkaline phosphatase and gamma-glutamyl-transferase in plasma were within reference levels. The mean of glutamate dehydrogenase (13.8 U/l) was slightly elevated in one flock. Mean liver concentrations of Zn (38.9 and 43.5 mg/kg ww) and Cu (111 and 87.5 mg/kg ww) in sheep flocks were higher compared to the respective roe deer populations (27.5 and 36.3 mg Zn/kg ww; 18.3 and 28.6 mg Cu/kg ww). This is supposed to be caused by differences in Cu and Zn metabolism in sheep and roe deer. Selenium deficiency was diagnosed in liver samples of both sheep flocks (0.21 and 0.23 mg/kg ww). There were neither significant differences compared to roe deer (0.21 and 0.27 mg Se/kg ww) nor differences depending on location. Correlations between plasma and liver concentrations of Cu, Zn and Se were not significant in sheep. Means of vitamin E in liver samples (30.6 and 41.8 mg/kg ww) were higher in roe deer populations. This may be caused by the opportunity of selective browsing for wild ruminants, which allows access to younger plants which are higher in vitamin E.     

Copper/zinc and copper/selenium ratios,and oxidative stress as biochemical markers in recurrent aphthous stomatitis

ProjectRecurrent aphthous stomatitis (RAS) is a common oral mucosal disorder characterized by recurrent, painful oral aphthae, and oxidative stress presumably contributes to its pathogenesis. The aim of this study is to scrutinize the relationship between oxidative stress and serum trace elements (copper, Cu; zinc, Zn; selenium, Se), and to evaluate the ratios of Cu/Zn and Cu/Se in this disorder.ProcedurePatients with RAS (n = 33) and age- and sex-matched healthy control subjects (n = 30) were enrolled in this study. Malondialdehyde (MDA) concentrations in plasma and the activities of superoxide dismutase (SOD1; CuZnSOD), glutathione peroxidase (GPx) and catalase (CAT) in erythrocyte were determined as spectrophotometric. Also, the levels of Se, Zn and Cu in serum were determined on flame and furnace atomic absorption spectrophotometer using Zeeman background correction.Results and conclusionsOxidative stress was confirmed by the significant elevation in plasma MDA, and by the significant decrease in CAT, SOD1, and GPx (p < 0.05). When compared to controls, Zn and Se levels were significantly lower in patients, whereas Cu levels was higher in RAS patients than those in controls (p < 0.05). In addition, the correlation results of this study were firstly shown that there were significant and positive correlations between Se–CAT, Se–GPx, and Cu–MDA parameters, but negative correlations between Se–Cu, Se–MDA, Cu–CAT, Cu–SOD1 and Cu–GPx parameters in RAS patients. Furthermore, the ratios of Cu/Zn and Cu/Se were significantly higher in the patients than the control subjects (p < 0.05). Our results indicated that lipid peroxidation associated with the imbalance of the trace elements seems to play a crucial role in the pathogenesis of RAS. Furthermore, the serum Cu/Zn and Cu/Se ratios may be used as biochemical markers in these patients.     

The importance of pyridoxine for the impact of the dietary selenium sources on redox balance,embryo development,and reproductive performance in gilts

This study aimed to determine the effects of dietary pyridoxine and selenium (Se) on embryo development, reproductive performance and redox system in gilts. Eighty-four gilts were fed one of five diets: CONT) basal diet; MSeB₆0) CONT + 0.3 mg/kg of Na-selenite; MSeB₆10) diet 2 + 10 mg/kg of HCl-pyridoxine; OSeB₆0) CONT + 0.3 mg/kg of Se-enriched yeast; and OSeB₆10) diet 4 + 10 mg/kg of HCl-pyridoxine. Blood samples were collected for long-term (each estrus and slaughter) and peri-estrus (fourth estrus d −4 to d +3) profiles. At slaughter (gestation d 30), organs and embryos were collected. For long-term and peri-estrus profiles, Se level and source affected (P < 0.01) blood Se concentration whereas B₆ level increased (P < 0.01) erythrocyte pyridoxal-5-phosphate concentration. A B₆ level (P < 0.05) effect was observed on long-term plasma Se-dependent glutathione peroxidase (Se-GPX) activity whereas peri-estrus Se-GPX was minimum on d −1 (P < 0.01). Selenium level increased sows’ organs and embryo Se concentration (P < 0.01). Selenium source tended to enhance embryo Se content (P = 0.06). Within-litter embryo Se content was increased by B₆ level (P < 0.01). Selenium level tended to affect Se-GPX and total GPX activities in organs mitochondria (P = 0.09 and 0.07, respectively). Selenium source affected kidney ATP synthesis (P = 0.05). In conclusion, B₆ level affected the Se-GPX activity on a long-term basis, whereas the basal level of Se was adequate during the peri-estrus period. Embryo quality was not improved by dietary Se, and B₆ impaired within-litter homogeneity.     

Release of ferulic acid from corn cobs by alkaline hydrolysis

The process of corn cobs alkaline hydrolysis to produce solutions with high hydroxy-cinnamic acids content was investigated. In particular the attention was focused on the solubilisation of ferulic acid (FA) and related compounds, mainly p-coumaric acid (p-CA). Although these compounds have applications as antioxidants, the purpose of this work was to obtain FA solutions that can be used as feedstock for the biotechnological production of vanillin in future studies. The effects of different concentrations of NaOH (0.2 ≤ C_a ≤ 2.0N) and solid/liquid ratios (0.028 ≤ S/L ≤ 0.168 g/g) on the solubilisation of FA versus time have been investigated at room temperature. Optimal hydrolysis conditions (C_a = 0.5N, S/L = 0.084 g/g after 6 h) ensured the production of hydrolysates with relatively high contents of both FA (1171 ± 34 mg/L) and p-coumaric acid (2156 ± 64 mg/L), which can be used in future studies for the microbial transformation into vanillin.     

Effects of zinc deficiency and zinc supplementation on homocysteine levels and related enzyme expression in rats

Methionine synthase (MS) and betaine-homocysteine methyltransferase (BHMT) are both zinc (Zn)-dependent methyltransferases and involved in the methylation of homocysteine. The objective of this study was to investigate the effects of dietary Zn supply on homocysteine levels and expression of the two enzymes in growing rats. Male weanling Sprague-Dawley rats were assigned randomly to four dietary groups (n = 8/group) for 3 weeks: Zn deficient (ZD; <1 mg Zn/kg); Zn control (ZC; 30 mg Zn/kg); Zn supplemented (ZS; 300 mg Zn/kg); pair fed (PF; 30 mg Zn/kg) to the ZD group. Serum and femur Zn concentrations were 83% and 58% lower in ZD, and 49% and 62% higher in ZS compared to ZC (P < 0.001), respectively. The ZD rats had lower feed intake (37%), body weight gains (45%), liver (43%) and kidney (31%) weights than those of ZC (P < 0.001), but these parameters in ZD were not significantly different from the PF controls. Serum homocysteine concentrations were 65% higher in ZD compared to PF (P < 0.05), and there was no significant difference in serum folate levels between ZD and PF groups. The mRNA expression of liver and kidney MS was 57% and 38% lower in ZD than PF (P < 0.001), respectively. Hepatic and renal BHMT mRNA levels were not altered in ZD compared to controls. The aforementioned measurements were not significantly different between ZS and ZC groups, except Zn levels. These results demonstrated that homocysteine homeostasis appeared to be disturbed by Zn deficiency but not Zn supplementation, and elevated serum homocysteine might be due to reduced expression of MS during Zn deficiency.     

Effects of selenium supplementation on selenium status of farmed fallow deer in outdoor pens

The study investigated the effects of selenium (Se) supplementation on Se status in farmed fallow deer. Fallow deer were housed on grass pasture and adapted to consume ∼200 g of pelleted grain daily. Animals were divided into two groups. One group received pelleted grain enriched with sodium selenate for 12 weeks (Se+ group, N = 10). Se intake for the first 7 weeks was 0.18 mg/kg dry matter (DM) and 0.32 mg/kg DM for the subsequent 5 weeks. The control group was fed pelleted grain without extra Se (Se− group, N = 9, 0.06-0.08 mg/kg DM). Blood samples were collected at the beginning and the end of the experiment. After the animals were slaughtered, tissue samples were collected for analysis of Se concentrations and Se-dependent glutathione peroxidase 1 (GPx1) activity. In addition, Se-independent α-glutathione-S-transferase (α-GST) activity was analyzed in liver tissue. Se supplementation significantly increased Se levels in plasma and in tissues as follows: liver > spleen > skeletal muscle > myocardium > kidney. Se supplementation also significantly increased GPx1 activity in tissues in the following order: liver > skeletal muscle > spleen = myocardium > kidneys. However, hepatic α-GST activity did not differ between Se+ and Se− groups. As expected, Se supplementation increased blood and tissue Se concentrations and GPx1 activity, which suggests a better antioxidant status. However, the activity of α-GST, an important Se-independent antioxidant enzyme, was not altered, presumably because GPx provided an adequate antioxidant capacity even though Se intake was low.     

Interaction between vitamin B6 and source of selenium on the response of the selenium-dependent glutathione peroxidase system to oxidative stress induced by oestrus in pubertal pig

This study aimed to assess the interaction between vitamin B₆ and selenium (Se) for the flow of Se towards the Se-dependent glutathione peroxidase (GPX) system in response to oxidative stress naturally induced by oestrus in a pubertal pig model. At first oestrus, forty-five gilts were randomly assigned to the experimental diets (n = 9/group): basal diet (CONT); CONT + 0.3 mg/kg of Na-selenite (MSeB₆0); MSeB₆0 + 10 mg/kg of HCl-B₆ (MSeB₆10); CONT + 0.3 mg/kg of Se-enriched yeast (OSeB₆0); and OSeB₆0 + 10 mg/kg of HCl-B₆ (OSeB₆10). Blood samples were collected at each oestrus (long-term profiles), and daily from day −4 to +3 (slaughter) of the fourth oestrus (peri-oestrus profiles) after which liver, kidneys, and ovaries were collected. For long-term profiles, CONT had lower blood Se than Se-supplemented gilts (p < 0.01) and OSe was higher than MSe (p < 0.01). Lower erythrocyte pyridoxal-5-phosphate was found in B₆0 than B₆10 (p < 0.01). No treatment effect was observed on GPX activity. For peri-oestrus profiles, treatment effects were similar to long-term profiles. Treatment effects on liver Se were similar to those for long-term blood Se profiles and OSe had higher renal Se concentrations than MSe gilts (p < 0.01). Gene expressions of GPX1, GPX3, GPX4, and selenocysteine lyase in liver and kidney were greatest in OSeB₆10 gilts (p < 0.05). These results suggest that dietary B₆ modulate the metabolic pathway of OSe towards the GPX system during the peri-oestrus period in pubertal pigs.     

Trace elements (copper,zinc, selenium and molybdenum) as markers in oral sub mucous fibrosis and oral squamous cell carcinoma

Oral cancer is a major cause of cancer morbidity and mortality worldwide and is prevalent in most areas where tobacco related practices are observed. Essential elements play a role in many biochemical reactions as a micro-source and there is growing evidence that their concentrations are altered on the onset and progress of malignant disease. In this study the levels of copper (Cu), zinc (Zn), selenium (Se) and molybdenum (Mo) in serum of patients with oral sub mucous fibrosis (OSMF) (n = 30) and oral squamous cell carcinoma (OSCC) (n = 30); were determined and the alterations of these critical parameters were analyzed in comparison with controls (n = 30) to identify predictors amongst these parameters for disease occurrence and progression. The serum Cu and Zn were established using Flame Atomic Absorption Spectrometry. Serum estimation of Se and Mo was done by graphite furnace atomic absorption spectrometry (GFAAS). Data analysis revealed a marked, progressive and significant increase in Cu levels in precancer (OSMF) and cancer (OSCC) groups as compared to the normal group. The level of Zn in serum was slightly elevated in OSMF and OSCC though not statistically significant. Cu/Zn ratio was slightly but not significantly elevated. Serum levels of Se and Mo were significantly decreased in the precancer and cancer groups as compared to the normals.     

The effect of age and gender on 38 chemical element contents in human iliac crest investigated by instrumental neutron activation analysis

ProjectTo understand the role of major, minor, and trace elements in the etiology of bone diseases including osteoporosis, it is necessary to determine the normal levels and age-related changes of bone chemical elements.ProcedureThe effect of age and gender on 38 chemical element contents in intact iliac crest of 84 apparently healthy 15–55 years old women (n=38) and men (n=46) was investigated by neutron activation analysis.ResultsMean values (M±SEM) for mass fraction (on dry weight basis) of Ca, Cl, Co, Fe, K, Mg, Mn, Na, P, Rb, Sr, and Zn for both female and male taken together were Ca – 169±3 g/kg, Cl – 1490±43 mg/kg, Co – 0.0073±0.0024 mg/kg, Fe – 177±24 mg/kg, K – 1820±79 mg/kg, Mg – 1840±48 mg/kg, Mn – 0.316±0.013 mg/kg, Na – 4970±87 mg/kg, P – 79.7±1.5 g/kg, Rb – 1.89±0.22 mg/kg, Sr – 312±15 mg/kg, and Zn – 65.9±3.4 mg/kg, respectively. The upper limit of mean contents of Cs, Eu, Hg, Sb, Sc, and Se were Cs≤0.09 mg/kg, Eu≤0.005 mg/kg, Hg≤0.005 mg/kg, Sb≤0.004 mg/kg, Sc≤0.001 mg/kg, and Se≤0.1 mg/kg, respectively. In all bone samples the contents of Ag, As, Au, Ba, Br, Cd, Ce, Cr, Gd, Hf, La, Lu, Nd, Sm, Ta, Tb, Th, U, Yb, and Zr were under detection limits.ConclusionsThe Ca, Mg, and P contents decrease with age, regardless of gender. Higher Ca, Mg, P, and Sr mass fractions as well as lower Fe content are typical of female iliac crest as compared to those in male bone.     

The relationship between fetal and maternal selenium concentrations in sheep and goats

In this study, biological samples (slaughterhouse material) were collected from 30 sheep and 36 goats and classified according to gestational stage into either early or late gestation. Samples consisted of allantoic fluid, amniotic fluid, fetal liver, fetal kidney, fetal thyroid gland, maternal plasma and liver to determine selenium (Se) concentrations throughout gestation. The Se concentrations in the allantoic fluid, fetal liver and kidney increased significantly (p < 0.01) during late gestation. Concurrently, the Se concentrations in amniotic fluid, maternal plasma and liver decreased significantly (p < 0.01) over time. Significant (p < 0.01) positive relationships were recorded between the age of the fetus and Se concentrations in the allantoic fluid (r = 0.57–0.75), fetal liver (r = 0.43–0.59) and kidney (r = 0.80–0.81) in both sheep and goats. A significant (p < 0.05) positive relationships were also recorded between the Se concentrations in the allantoic fluid and fetal liver (r = 0.35–0.37), the maternal plasma and liver Se concentrations (r = 0.37–0.57) between sheep and goats. A significant (p < 0.05) negative correlation was recorded between the Se concentrations in the allantoic fluid with maternal plasma of sheep (r = −0.41) as well as between the fetal liver and maternal liver Se (r = −0.22 to 0.50) and a negative correlation (r = −0.42 to 0.43) (p < 0.01) between Se concentrations in the fetal liver and amniotic fluid in both sheep and goats, respectively. Se concentration in the fetal liver was significantly (p < 0.01) higher than that of the kidney and thyroid. In the thyroid gland no morphological differences were noted. Strong fetal–maternal relationships in Se concentration were evident throughout the gestational period and dams seem to sacrifice Se levels in order to maintain that in the fetus. Se concentrations in the amniotic and allantoic fluids could be used as a possible indicator of the Se status of the fetus throughout gestation.     

Toxic and essential elements in children's blood (<6 years) from Kinshasa,DRC (the Democratic Republic of Congo)

In this study we determined the concentration of 9 trace elements (As, Cd, Cu, Hg, Mn, Mo, Pb, Se and Zn) in whole blood of children (n = 100, 64 girls, 36 boys and median age: 36 months) using inductively coupled plasma mass spectrometry (ICP-MS). The proportion of children potentially deficient in essential elements or poisoned by toxic elements was evaluated. The aging effects on the concentration of these elements were also investigated. The median values were 3.17 μg/L (As), 0.15 μg/L (Cd), 1.1 mg/L (Cu), 2.1 μg/L (Hg), 10.4 μg/L (Mn), 17.7 μg/L (Mo), 8.7 μg/dL (Pb), 10.7 μg/L (Se) and 5.0 mg/L (Zn). The concentration of many elements (As, Cd, Hg, Mn, Pb and Zn) showed significant age variations but not sex influence. Regarding levels of the essential elements (Cu, Mn, Mo, Se and Zn), B-Cu, B-Mn, B-Se and B-Zn were in the normal range, whereas exceeded levels were observed for B-Mo. None of these children was deficient in essential elements. Except B-Cd, all toxic elements showed exceeded blood levels. The proportion of children potentially poisoned by toxic elements varies from 10% (n = 10) to 95% (n = 95) and depends on toxic element: 95% for As, 10% for Hg and 35% for Pb. The main health concerns emerging from this study are the high As, Hg and Pb exposures of the Kinshasan children requiring further documentation, corrective actions and the implementation of appropriate regulations.     

Quantitative analysis of RU38486 (mifepristone) by HPLC triple quadrupole mass spectrometry

A sensitive liquid chromatography–mass spectrometric method was validated for the quantification of RU38486 (mifepristone) in human and murine plasma. The analyte and internal standard (alfaxolone) were extracted by liquid–liquid extraction with diethyl ether, resolved on a C18 column using gradient elution with methanol and ammonium acetate and detected after positive electrospray ionization (m/z 430 → 372; m/z 333 → 297, respectively). Quantification was linear over the range 0.5–500 ng (r² > 0.997), precise and accurate (intra-assay RSD ≤ 7.2%, RME ≤ 8.2%; inter-assay RSD ≤ 15.7% RME ≤ 10.2%). The limit of quantification (LOQ) was 50 pg injected on column, permitting reproducible analysis of RU38486 in small volumes of plasma.     

Selenium uptake,speciation and stressed response of Nicotiana tabacum L.

The research on the function and mechanism of selenium (Se) is of great significance for the development of Se-enriched agricultural products. In this paper, uptake, speciation distribution, the effects on the flue-cured tobacco growth and antioxidant system of Se at different levels (0–22.2 mg Se kg⁻¹) were studied through a pot experiment, aiming to clarify flue-cured tobacco's response to Se stress and the relationship between Se speciation and antioxidant system. The results showed that the leaf area and number, the biomass and the chlorophyll content reached the maximum at 4.4 mg kg⁻¹ of Se treatment. Selenium at low levels (≤4.4 mg kg⁻¹) stimulated the growth of flue-cured tobacco by elevating the capability of antioxidant stress and reducing the malondialdehyde (MDA) content to 0.6–0.8 times of that of the control. However, high Se levels (≥11.1 mg kg⁻¹) depressed the capability of antioxidant stress and raised the MDA content to 1.5-fold of that of the control, and meanwhile the biomass of the aboveground parts and underground parts declined notably. The Se content in different parts of flue-cured tobacco significantly increased with the growth of Se levels. The range of Se content in roots, leaves and stems at 2.2–22.2 mg kg⁻¹ of Se treatment were 16.7–58.6 mg kg⁻¹, 2.6–37.3 mg kg⁻¹ and 2.2–10.3 mg kg⁻¹, respectively. According to the detection of different Se speciation, only selenocysteine (SeCys) was detectable in leaves at 2.2 mg kg⁻¹ Se treatment; SeCys, selenite [Se(IV)]and selenate [Se(VI)] were detected in flue-cured tobacco leaves at Se treatment (≥4.4 mg kg⁻¹), which accounted for 4.6–10%, 9–18.7% and 71–86% respectively; SeCys, selenomethionine (SeMet) and Se(IV) were detected in roots, and organic selenium(66–84%) was the main Se species at Se ≤ 11.1 mg kg⁻¹ treatment; four Se species [SeCys, SeMet, Se(IV) and Se(VI)] were detected in flue-cured tobacco roots, and the main Se species was inorganic Se (60%) at 22.2 mg kg⁻¹ Se treatment. That was to say, the percentage of organic Se species (SeCys and SeMet in flue-cured tobacco leaves and root) declined, whereas the ratio of inorganic Se species [Se(IV) and Se(VI)] increased with the growth of Se levels. The correlation analysis showed that the superoxide dismutase (SOD) activity as well as the glutathione (GSH) and MDA contents were positively correlated with the Se(IV) and Se(VI) contents at P < 0.01 and excessive inorganic Se might destruct the reactive oxygen species (ROS) balance and enhance the MDA content, thus causing damage to the plant growth. In a word, the present study suggested that the ratio of inorganic Se [Se(IV) and Se(VI)] was closely related with the growth and the antioxidant capacity of flue-cured tobacco and the excessive application of Se led to the higher proportion of inorganic Se and poorer antioxidant capacity, which ultimately inhibited the growth of flue-cured tobacco.     

Relative bioavailabilities of organic zinc sources with different chelation strengths for broilers fed diets with low or high phytate content

A six-day experiment was conducted to estimate the relative bioavailability values (RBV) of zinc (Zn) in three organic sources (oZn) with different chelation strengths compared to inorganic ZnSO₄ (iZn) for broilers fed a low or high phytate diet. A total of 1080, one-d-old male broiler chicks were randomly assigned to one of 18 dietary treatments (six replicates cages of ten chicks per cage) in a completely randomized design involving a 2 × 2 × 4 factorial arrangement with two levels of added phytate (0 or 10 g phytate as sodium phytate/kg), two levels of added Zn (30 or 60 mg/kg) and four Zn sources (iZn and three oZn sources) plus one low and one high phytate control treatments without Zn addition. The three oZn sources consisted of (1) Zn amino acid with weak chelation strength (ZnAA-L, formation quotient Q_f = 6.6, containing 119 g Zn/kg), (2) Zn proteinate with moderate chelation strength (ZnPRO-M, Q_f = 30.7, containing 133 g Zn/kg) or (3) Zn proteinate with strong chelation strength (ZnPRO-H, Q_f = 944.0, containing 186 g Zn/kg). Chicks were harvested at 6 days of age and pancreas metallothionein (MT) mRNA expression was used to estimate Zn RBV. Pancreas MT mRNA expression increased (P<0.01) as dietary Zn level increased. Chicks fed high phytate diets had lower (P<0.05) MT mRNA expression than chicks fed low phytate diets. Based on multiple linear regression slope ratios with ZnSO₄ set at 1.00, the RBV of ZnAA-L, ZnPRO-M and ZnPRO-H were 1.01, 1.28 and 0.70, respectively, for low phytate diets, and 1.05, 1.39 and 0.92, respectively, for high phytate diets. The slope for the oZn source with moderate chelation strength differed (P<0.05) from iZn and the other two oZn sources. The RBV of ZnAA-L, ZnPRO-M and ZnPRO-H under the high phytate diet increased by 0.04, 0.11 and 0.22, respectively, compared to those under the low phytate diet. Results indicate that the oZn sources with moderate or strong chelation strength offer partial or complete resistance to interference from high dietary phytate during digestion; and the oZn with moderate chelation strength had a greater RBV with both low and high phytate diets than iZn or oZn sources with weak or strong chelation strength.     

Elements of margin of safety,toxicity and action of sodium selenite in a lipopolysaccharide rat model

ProjectBoth septic shock and sodium selenite (Na₂SeO₃) lead to multiple organ failure through oxidation. Na₂SeO₃ has direct oxidant effects above the nutritional level and indirect anti-oxidant properties.In a lipopolysaccharide (LPS) rat model we assessed margin of safety, toxicity and beneficial effect of pentahydrate Na₂SeO₃ (5H₂O·Na₂SeO₃) at oxidant doses.ProcedureIn a three-step study on 204 rats we: (i) observed toxic effects of Na₂SeO₃ injected intraperitoneously (IP) and determined its Minimum Dose Without Toxic effect (MDWT) 0.25–0.35 mg/kg selenium (Se) content; (ii) injected IP LPS at 70% lethal dose (LD) followed, or not, one hour later by IP Na₂SeO₃ at MDWT and (iii) by doses > MDWT. At 48 h, in survivors, we measured plasma creatinine, lactate, aspartate and alanine aminotransferase (AST, ALT), nitric oxide (NO) and Se concentrations.Results(i) Na₂SeO₃ alone did not increase NO and lactate. Encephalopathy appeared at 1 mg Se/kg. Creatinine increased at 1–1.75 mg Se/kg, AST, ALT at 3–4.5 mg Se/kg, and the minimum LD was 3 mg Se/kg. (ii) Mortality after LPS was 37/50 (74%, [62–86%]) vs. 20/30 (67%, [50–84%]) when followed by Na₂SeO₃ at MDWT (p = 0.483) with a decreased in NO (−31%, p = 0.038) a trend for lactate decrease (−19%, p = 0.068) and an increased Se in plasma of survivals. (iii) All rats died at doses ≥0.6 mg/kg (p < 0.001).ConclusionMechanisms of LPS and Na₂SeO₃ toxicity differ (i.e. NO, lactate). In septic shock 5H₂O·Na₂SeO₃ toxicity increased, margin of safety decrease, but IP administration of dose considered as oxidant of 5H₂O·Na₂SeO₃ showed beneficial effects.     

Cellular and nephrotoxicity of selenium species

ProjectBeside its useful functions at very low concentrations, selenium including supplementary Se sources pose a potential toxicological risk. The toxicity of selenium species was tested in HaCaT cell culture and related nephrotoxicity in mice.ProcedureThe apoptotic shrinkage and necrotic expansion of cells were measured by time-lapse image microscopy. Acute nephrotoxicity was estimated upon administration of various selenium species to mice for two weeks. To confirm or to refute the accumulation of Se in the kidney and its potential chronic effect, Se concentration in kidney tissue and histopathlology were tested.ResultsThe comparison of selenium species showed that organic lactomicroSe did not affect cell growth at 5 ppm, but inorganic nanoSe severely hampered it at lower concentration (1 ppm). The in vivo Se treatment (0.5, 5, 50 ppm, corresponding to 4, 40 and 400 μg/kg) was misleading as it did neither affect the outward appearance nor the weight of the kidney. Se accumulation was observed after selenate, selenite, SelPlex, selenite and nanoSe administration, while lactomicroSe caused no traceable accumulation. In vivo, ex vivo and in vitro experiments reflected this order of selenium toxicity: selenate > selenite > SelPlex = nanoSe > lactomicroSe.ConclusionWithin the tested species lactomicroSe was the only non-nephrotoxic selenium source recommended for nutritional Se supplementation.     

Pharmacological studies on the sedative-hypnotic effect of Semen Ziziphi spinosae (Suanzaoren) and Radix et Rhizoma Salviae miltiorrhizae (Danshen) extracts and the synergistic effect of their combinations

Semen Ziziphi spinosae (Suanzaoren in China) and Radix et Rhizoma Salviae miltiorrhizae (Danshen in China) are conventional herbal drugs in traditional Chinese medicine and have been used widely for the treatment of insomnia. In the present study, the sedative-hypnotic activity of the active fractions extracted from Suanzaoren and Danshen were studied using the method of pentobarbital-induced sleep in the mouse model. Qualitative analysis of the standardized extracts was carried out by HPLC-DAD. The results showed that the water extract of Suanzaoren (SWE) (400 and 800 mg/kg body wt.) and the ether extract of Danshen (DTT) (300 and 600 mg/kg body wt.) can shorten sleep latency significantly, increase sleeping time and prolong movement convalescence time induced by sodium pentobarbital (55 mg/kg body wt.) administration in mice. Furthermore, the combination of SWE and DTT showed significant synergistic effect (p<0.05) in decreasing sleep latency and increasing sleeping time, but not in prolonging the movement convalescence time, which might be helpful for energy recovery in the treatment of insomnia. The results suggest that SWE, DTT, and the combination of SWE and DTT possess significant sedative-hypnotic activity, which supports the popular use of Suanzaoren and Danshen for treatment of insomnia and provide the basis for new drug discovery. Furthermore, the results demonstrate that the combination of SWE and DTT may be preferable for the treatment of insomnia.     

Effect of suplemental nitrogen source and feeding frequency on nutrient supply to lambs fed a kikuyu grass (Pennisetum clandestinum) hay-based diet

Eight castrated male lambs (35 ± 4 kg live weight), fed a basal diet of kikuyu grass hay, were used in a replicated 4 × 4 Latin Square experiment with a 2 × 2 factorial arrangement of treatments to evaluate the effect of supplemental feeding frequency and source of rumen degradable N on intake, digestibility, ruminal fermentation, and microbial protein yield. Treatments were supplementation with cassava meal plus calcium caseinate or cassava meal plus urea offered at a rate of 7 g/kg live weight daily in one or two meals per day. Lambs were fed twice daily in such manner to allow ad libitum comsumption of forage. There was significant feeding frequency by N source interaction on variables of intake. In general, intake of feed components was higher (P ≤ 0.05) by lambs offered the caseinate-supplement twice daily over intake observed in lambs given the others diet treatments. Digestibility of feed components was neither affected by supplemental N source (DM, P = 0.541; OM, P = 0.585; NDF, P = 0.828) nor by feeding frequency (DM, P = 0.122; OM, P = 0.175; NDF, P = 0.591). Urinary excretion of N increased (P ≤ 0.05) in lambs supplemented twice daily whereas N retention was similar for all treatments (N source, P = 0.748; feeding frequency, P = 0.418). Microbial protein entering into the small intestine was affected by the interaction between feeding frequency and N source such as an increasing (P < 0.10) in this variable was observed when lambs received the caseinate but not the urea supplement twice daily. Efficiency of microbial protein synthesis, however, was not affected by treatments (N source, P = 0.588; feeding frequency, P = 0.334). Rumen pH averaged 6.70 and it was neither affected by N source (P = 0.827) nor by feeding frequency (P = 0.740). Ruminal concentration of ammonia N was not affected by feeding frequency (P = 0.144) while it increased (P < 0.05) when urea rather than caseinate was the supplemental N source (mean of 7.61 mg/dl vs. 6.00 mg/dl). Concentration of sugars in rumen fluid was higher (P ≤ 0.05) in lambs supplemented once a day compared to twice daily (mean of 49.4 mg/dl vs. 34.4 mg/dl) for both N sources. A significant (P ≤ 0.05) N source by feeding frequency interaction effect was observed for ruminal concentrations of α-amino N compounds. In urea treatment α-amino N concentration increased (P ≤ 0.05) in lambs receiving the supplement twice daily compared to once a day (mean of 4.59 mg/dl vs. 3.70 mg/dl) while in caseinate treatment it was higher (P ≤ 0.05) in lambs offered the supplement in one meal per day compared to twice daily (mean of 5.29 mg/dl vs. 4.07 mg/dl). In conclusion, for ruminants fed a tropical grass-based diet, starch-rich supplement containing non-protein N as N source may be offered only once a day whereas the supply of nutrients may be improved if degradable true protein is included as N source and supplement is offered in two meals per day.