Sources and levels of copper affect liver copper profile, intestinal morphology and cecal microbiota population of broiler chickens fed wheat-soybean meal diets.

Super dosing copper (Cu) has long been used as an alternative to antibiotic growth-promoters in broiler chickens' diet to improve gut health. This study was designed to compare nutritional and growth-promoting levels of Cu hydroxychloride (CH) with CuSO4 on gut health bio-markers and liver mineral profile of broiler chickens. Ross 308 chicks (n = 864) were randomly assigned to eight treatments, as basal diet containing no supplemental Cu; the basal diet with 15 or 200 mg/kg Cu as CuSO4; or 15, 50, 100, 150 or 200 mg/kg Cu from CH. The highest liver Cu content was observed in birds fed the diets with 200 mg/kg CuSO4 (P < 0.01). Serum FITC-d concentration as the leaky gut marker, and liver malondialdehyde concentration were not affected. Copper level or source had no effect on cecal short chain fatty acid and the mRNA expression of five jejunal genes involved in gut integrity. Negative linear responses of Cu were observed on Lactobacillus (P = 0.032), Bacteroides (P = 0.033), and Enterobacteriaceae (P = 0.028) counts. The jejunal villus height increased in birds fed CH at 200 and 100 mg/kg (P < 0.05). Increasing Cu levels, linearly and quadratically (P < 0.001), increased Cu excretion.

Chronic tribasic copper chloride exposure induces rat liver damage by disrupting the mitophagy and apoptosis pathways.

Despite the fact that copper (Cu) is a vital micronutrient to maintain body function, high doses of Cu through environmental exposure damage various organs, especially the liver, which is the main metabolic organ. To investigate the influence of long-term Cu-induced toxicity on mitophagy and apoptosis in rat liver, 96 seven-month-old male Sprague-Dawley rats were fed TBCC for 24 weeks. The results revealed that exposure to high Cu concentrations could promote oxidative stress liver injury by increasing the hepatic function index (ALT, AST and ALP) and MDA content, while reducing the activity of antioxidant enzymes (T-SOD, GSH-Px and CAT) related to oxidative stress. Consistent with histopathological observations, proper dietary Cu (15-60 mg/kg) could improve antioxidant stress levels and induce a dose-dependent increase in the mRNA expression of mitophagy-related genes, whereas a high Cu concentration (120 mg/kg) could cause severe liver impairment and ultrastructural changes and a reduction in mitophagosomes, accompanied by downregulation of Atg5, Beclin1, Pink1, Parkin, NIX, P62 and LC3B. The expression of apoptosis-related genes (Bax, Bax/Bcl-2, Caspase3, Cytc and p53) and proteins (Caspase3 and p53) was upregulated with the addition of dietary Cu. The results demonstrated that an appropriate dose of TBCC could improve liver function by promoting mitophagy and Cu enzymes that play antioxidative roles, while the accumulation of excess Cu could induce liver lesions by enhancing apoptosis and inhibiting mitophagy pathways.

Chronic Copper Exposure Induces Hypospermatogenesis in Mice by Increasing Apoptosis Without Affecting Testosterone Secretion.

Chronic copper exposure impaired spermatogenesis in adult male mice. The aim of this study was to determine whether chronic copper exposure can induce apoptosis of testicular cell and hypospermatogenesis via disturbing testosterone synthesis in adult male mice. In the present study, sixty CD-1 male mice were randomly divided into four groups, and were continuously administered for 8 weeks by oral gavage with copper sulfate at a dose of 0, 25, 100, and 150 mg/kg/day, respectively. We determined the content of serum and testicular copper, testicular coefficient, testicular histopathology, sperm count and motility, the mRNA and protein levels of Caspase-3, Bax, and Bcl-2, Leydig cell count, testosterone content, testosterone synthetase, and testosterone synthesis-related genes. The results showed that the copper levels in serum increased in a dose-dependent manner, and the copper levels in testes were significantly related to serum copper levels. Male mice given copper sulfate 100 and 150 dosage groups showed significant decreased in sperm motility and sperm number as well as increased in testes damage, and there was no significant change in testicular coefficient in the four groups. The mRNA levels of Bcl-2 decreased and Caspase-3 increased in 150 dosage group, and Bax increased in two higher dosage groups. Meanwhile, Caspase-3 and Bax proteins increased in 150 dosage group, and Bcl-2 protein decreased in three copper treatment groups. Nevertheless, there were no differences on the levels of testosterone content and testosterone synthetase of 3ß-HSD, 17ß-HSD, 17α-Hyd, and 20α-Hyd, mRNA levels of Cyp11a1, Cyp17a1, and Star, and quantity of Leydig cells in four groups. Overall, these data showed that chronic copper exposure led to copper residues in the testes, and the doses of 100 and 150 mg/kg/day copper sulfate may induce hypospermatogenesis by increasing apoptosis without affecting testosterone secretion.

Noninvasive rapid bacteria-killing and acceleration of wound healing through photothermal/photodynamic/copper ion synergistic action of a hybrid hydrogel.

Bacterial infection often delays healing of wounded tissues and so it is essential to improve the antibacterial efficiency in situ. In this work, a hybrid hydrogel composed of 3-(trimethoxysilyl)propyl methacrylate (MPS, 97%) and mesoporous silica (mSiO2) modified CuS nanoparticles (NPs) is synthesized by radical polymerization. The materials possess excellent and controllable photothermal and photodynamic properties under 808 nm near-infrared (NIR) light irradiation as well as an antibacterial efficacy of 99.80% and 99.94% against Staphylococcus aureus and Escherichia coli within 10 min, respectively. The excellent performance stems from the combined effects of hyperthermia, radical oxygen species, and released copper ions produced during NIR irradiation of CuS NPs. Moreover, the released copper ions stimulate fibroblast proliferation and angiogenesis and the intrinsic volume transition of the hydrogel composed of N-isopropylacrylamide (NIPAAm) and acrylamide (AAm) controls the release rate of copper ions during NIR light irradiation leading to both antibacterial effects and skin tissue regeneration.

Disposable luciferase-based microfluidic chip for rapid assay of water pollution.

In the present study, we demonstrate the use of a disposable luciferase-based microfluidic bioassay chip for environmental monitoring and methods for fabrication. The designed microfluidic system includes a chamber with immobilized enzymes of bioluminescent bacteria Photobacterium leiognathi and Vibrio fischeri and their substrates, which dissolve after the introduction of the water sample and thus activate bioluminescent reactions. Limits of detection for copper (II) sulfate, 1,3-dihydroxybenzene and 1,4-benzoquinone for the proposed microfluidic biosensor measured 3 µM, 15 mM, and 2 µM respectively, and these values are higher or close to the level of conventional environmental biosensors based on lyophilized bacteria. Approaches for entrapment of enzymes on poly(methyl methacrylate) (PMMA) plates using a gelatin scaffold and solvent bonding of PMMA chip plates under room temperature were suggested. The proposed microfluidic system may be used with some available luminometers and future portable luminescence readers.

Impact of copper sulfate application at an urban Brazilian reservoir: A geostatistical and ecotoxicological approach.

A landscape ecotoxicology approach was used to assess the spatial distribution of copper in the recent bottom sediment (surficial sediment) of a Brazilian subtropical reservoir (the Guarapiranga reservoir) and its potential ecotoxicological impacts on the reservoir ecosystem and the local society. We discuss the policies and procedures that have been employed for the management of this reservoir over the past four decades. Spatial heterogeneity in the reservoir was evaluated by means of sampling design and statistical analysis based on kriging spatial interpolation. The sediment copper concentrations have been converted into qualitative categories in order to interpret the reservoir quality and the impacts of management policies. This conversion followed the Canadian Water Framework Directive (WFD) ecotoxicological concentration levels approach, employing sediment quality guidelines (SQGs). The SQG values were applied as the copper concentration thresholds for quantitative-qualitative conversion of data for the surficial sediment of the Guarapiranga. The SQGs used were as follows: a) interim sediment quality guideline (ISQG), b) probable effect level (PEL), and c) regional reference value (RRV). The quantitative results showed that the spatial distribution of copper in the recent bottom sediment reflected the reservoir's management policy and the copper application protocol, and that the copper concentrations varied considerably, ranging from virtually-zero to in excess of 3gcopper/kgds. The qualitative results demonstrated that the recent bottom sediment was predominantly in a bad or very bad condition, and could therefore have impacts on the local society and the ecosystem. It could be concluded that the management policy for this reservoir was mainly determined by the desire to minimize short-term costs, disregarding long-term socioeconomic and environmental consequences.

Modification of chitosan/calcium alginate/Fe3O4 hydrogel microsphere for enhancement of Cu(II) adsorption.

Copper in drinking water causes a significant environmental problem. Composite material based on alginate hydrogel has been gaining attention in the field of Cu(II) adsorption. However, alginate-based hydrogel exhibits poor mechanical property and relative low adsorption capacity which limit their application. The present study is devoted to the modification of chitosan/calcium alginate/Fe3O4 (CAF) hydrogel microsphere by NaOH solution for enhancement of Cu(II) adsorption. Results reveal that modification of CAF via NaOH solution significantly improves the mechanical strength and Cu2+ adsorption capacity of pristine materials. FTIR and XRD analysis confirms that CAF and newly prepared materials (NACAF) are successfully prepared. SEM and EDX are employed to analyze the surface morphology and elemental composition, respectively, both before and after their loading with Cu2+. XPS study demonstrates adsorption mechanism is based on chelation and ion-exchange. Compressive stress-strain curves demonstrate NACAF has better mechanical performance than CAF. The adsorption kinetics of the two adsorbents follow a pseudo-second-order model. The equilibrium data were best described by Langmuir isotherm model, and the estimated maximum equilibrium sorption capacity, q m,is 261.31 mg/g for the NACAF, which is larger than that of CAF (145.39 mg/g). Hence, NACAF shows excellent mechanical strength and high sorption capacity for Cu2+. It has great potential for Cu(II) removal in aqueous solutions.

Relative bioavailability of copper in tribasic copper chloride to copper in copper sulfate for laying hens based on egg yolk and feather copper concentrations.

This experiment was conducted to determine the relative bioavailability (RBV) of Cu in tribasic copper chloride (TBCC) to Cu in copper sulfate (monohydrate form; CuSO4·H2O) for layer diets based on egg yolk and feather Cu concentrations. A total of 252, 72-wk-old Hy-Line Brown laying hens were allotted to 1 of 7 treatments with 6 replicates consisting of 6 hens per replicate in a completely randomized design. Hens were fed corn-soybean meal-based basal diets supplemented with 0 (basal), 100, 200, or 300 mg/kg Cu from CuSO4 or TBCC for 4 wk. Results indicated that egg production, egg weight, and egg mass were not affected by dietary treatments. However, increasing inclusion levels of Cu in diets from CuSO4 decreased (P < 0.05) feed conversion ratio (FCR), whereas increasing inclusion levels of Cu in diets from TBCC did not affect FCR, indicating significant interaction (P < 0.05). Increasing inclusion levels of Cu from TBCC or CuSO4 increased (P < 0.05) Cu concentrations of egg yolk and feathers. Feather Cu concentrations were greater (P < 0.01) for hens fed diets containing CuSO4 than for hens fed diets containing TBCC. The values for the RBV of Cu in TBCC to Cu in CuSO4 based on log10 transformed egg yolk and feather Cu concentrations were 107.4% and 69.5%, respectively. These values for the RBV of Cu in TBCC did not differ from Cu in CuSO4 (100%). The RBV measured in egg yolk did not differ from the RBV measured in feather. In conclusion, the RBV of Cu in TBCC to Cu in CuSO4 can be determined using Cu concentrations of egg yolk and feathers although the values depend largely on target tissues of laying hens. For a practical application, however, the RBV value of Cu in TBCC to Cu in CuSO4 could be 88.5% when the RBV values determined using egg yolk and feather Cu concentrations were averaged.

Effects of two copper compounds on Microcystis aeruginosa cell density, membrane integrity, and microcystin release.

Microcystin release following Microcystis aeruginosa cell lysis after copper-based algaecide treatment is often cited as a concern leading to restricted use of algaecide in restoration of natural water resources. To examine this concern, bench-scale experiments were conducted to study responses of M. aeruginosa to 8-day copper exposures as copper sulfate and copper-ethanolamine (Cu-EA). M. aeruginosa UTEX 2385 was cultured in BG11 medium to cell density of 10(6)cells/mL with total and extracellular microcystin of 93 and 53µg/L, respectively. Exposures of copper concentration ranged from 40 to 1000µgCu/L. Cell membrane integrity was indicated by erythrosine B. In the end of experiment, total microcystin and cell density in untreated control (313µg/L and 10(7)cells/mL) was 3.3 and 10 times greater than pretreatment value, respectively. Minimum amount of copper required to reduce M. aeruginosa population within 8 days was 160µgCu/L as copper sulfate and 80µgCu/L as Cu-EA, where total and extracellular microcystin concentrations (47 and 44µg/L for copper sulfate; 56 and 44µg/L for Cu-EA) were degraded with degradation rate coefficient 0.1 day(-1) and were less than pretreatment values. Given a copper concentration at 80µgCu/L as Cu-EA, M. aeruginosa cells were intact and less microcystin were released compared to treatments at 160-1000µgCu/L, where lysed cells and relatively greater microcystin release were observed. Based on the laboratory results, a minimum amount of copper required for reducing M. aeruginosa population could decrease total microcystin concentration and not compromise cells and minimize microcystin release.

Exploiting external reflection FTIR spectroscopy for the in-situ identification of pigments and binders in illuminated manuscripts. Brochantite and posnjakite as a case study.

In the present work, the use of portable instrumentation allowing in-situ reflection FTIR analyses is exploited to identify the coloring matters of northern-Italian illuminations dating to the XVI century. In order to build a database of spectra, reference paint samples were prepared spreading the pigments on parchment with two different binders, i.e. gum arabic and egg white, used in antiquity. Pigments for the database were chosen considering their use in the Middle Ages and in the Renaissance and their response in the mid- and near-IR region. The reflection FTIR spectra obtained resulted to be dominated by the specular reflection component, allowing the use of the Kramers-Kronig transform to convert them to the more conventional absorbance FTIR spectra. Several pigments could thus be identified in ancient illuminations, even if some green details showed a spectral pattern different with respect to the most common commercial green pigments of the database. Therefore, in addition, basic copper sulfates brochantite and posnjakite were synthesized and characterized. In three green details, posnjakite was identified, both as a pure compound and together with malachite.

Effect of different glycation agents on Cu(II) binding to human serum albumin, studied by liquid chromatography, nitrogen microwave-plasma atomic-emission spectrometry, inductively-coupled-plasma mass spectrometry, and high-resolution molecular-mass spectrometry.

The ability of human serum albumin to capture unbound copper under different clinical conditions is an important variable potentially affecting homeostasis of this element. Here, we propose a simple procedure based on size-exclusion chromatography with on-line UV and nitrogen microwave-plasma atomic-emission spectrometry (MP-AES) for quantitative evaluation of Cu(II) binding to HSA upon its glycation in vitro. The Cu-to-protein molar ratio for non-glycated albumin was 0.98 ± 0.09; for HSA modified with glyoxal (GO), methylglyoxal (MGO), oxoacetic acid (GA), and glucose (Glc), the ratios were 1.30 ± 0.22, 0.72 ± 0.14, 0.50 ± 0.06, and 0.95 ± 0.12, respectively. The results were confirmed by using ICP-MS as an alternative detection system. A reduced ability of glycated protein to coordinate Cu(II) was associated with alteration of the N-terminal metal-binding site during incubation with MGO and GA. In contrast, glycation with GO seemed to generate new binding sites as a result of tertiary structural changes in HSA. Capillary reversed-phase liquid chromatography with electrospray-ionization quadrupole-time-of-flight tandem mass spectrometry enabled detection and identification of Cu(II) coordinated to the N-terminal metal-binding site (Cu(II)-DAHK) in all tryptic digests analyzed. This is the first report confirming Cu(II)-DAHK species in HSA by means of high-resolution tandem mass spectrometry, and the first report on the use of MP-AES in combination with chromatographic separation.

Effects of copper sulfate on seedlings of Prosopis pubescens (screwbean mesquite).

Phytoextraction is an established method of removal of heavy metals from contaminated soils worldwide. Phytoextraction is most efficient if local plants are used in the contaminated site. We propose that Prosopis pubescens (Screw bean mesquite) would be a successful phytoextractor of copper in our local soils. In order to determine the feasibility of using Screw bean mesquite, we utilized inductively-coupled plasma-optical emission spectroscopy (ICP-OES) and elemental analysis to observe the uptake of copper and the effects on macro and micro nutrients within laboratory-grown seedlings. We have previously shown that P. pubescens is a hyperaccumulator of copper in soil-grown seedlings. Light and transmission electron microscopy demonstrated death of root cells and ultrastructural changes due to the presence of copper from 50 mg/L - 600 mg/L. Ultrastructural changes included plasmolysis, starch accumulation, increased vacuolation and swollen chloroplasts with disarranged thylakoid membranes in cotyledons. Inductively coupled plasma-optical emission spectroscopy analyses of macro- and micro-nutrients revealed that the presence of copper sulfate in the growth medium of Petri-dish grown Prosopis pubescens seedlings resulted in dramatic decreases of magnesium, potassium and phosphorus. At 500-600 mg/L of copper sulfate, a substantial increase of sulfur was present in roots.

The potential toxicity of copper nanoparticles and copper sulphate on juvenile Epinephelus coioides.

Copper nanoparticles (Cu-NPs) were widely used in various industrial and commercial applications. In this study the effects of Cu-NPs and soluble Cu were investigated on juvenile Epinephelus coioides. The fish were exposed in triplicate to control, 20 or 100µgCuL(-1) as either copper sulphate (CuSO4) or Cu-NPs in a semi-static aqueous culture for 25 days. The growth parameters were significantly lower at 100µgCuL(-1) as CuSO4 or Cu-NPs treatment compared to control. Time-dependent Cu accumulation in all tissues increased with increasing the Cu dose. The percentage of total Cu found in remaining tissues (head, bones, fins, etc.) decreased more in the CuSO4 than Cu-NPs treatment after 25 days, but increased in all other tissues (especially in liver). Compared with the control, either Cu-NPs or CuSO4 induced higher malonaldehyde concentration in tissues by overwhelming total superoxide dismutase activity, total glutathione concentration and Na(+)/K(+)-ATPase activity, but the opposite results were recorded for the brain. With increasing the CuSO4 or Cu-NPs dose, apoptosis was exacerbated in the liver and gills, more so by CuSO4 than Cu-NPs. Overall, these findings showed that Cu-NPs had the toxic effects similar to dissolved Cu; hence, Cu-NPs need to be included in the assessment of toxicological impacts in the aquatic environment.

Direct separation of faradaic and double layer charging current in potential step voltammetry.

Double layer charging current in electrochemical systems has been a challenging problem in the last several decades because it causes interference to the accurate measurement of faradaic current. A method for extracting faradaic current and double layer charging current directly from the measured total current in potential step voltammetry is developed by using iterative target transformation factor analysis (ITTFA). The method constructs initial target vectors based on the theoretical formulae of faradaic and charging current, and then calculates the weights of faradaic and charging current in the measured signal via the iterative transformation of the initial vectors. Therefore, the two currents in one experiment can be obtained simultaneously without any assumption. The potential step voltammetric signals of potassium ferricyanide, copper sulfate and paracetamol were analyzed with the proposed method. The results show that the shape of the obtained voltammogram is an ideal sigmoid curve with horizontal straight baseline and plateaus, and the intensity of the signal is greatly enhanced. Therefore, the method provides a new way to measure the pure faradic current in the potential step voltammetric experiment, and may provide an alternative for improving the sensitivity of quantitative analysis.

Efeitos do tratamento com formalina e sulfato de cobre sobre os parâmetros hematológicos e parasitos monogenéticos em juvenis de Hemigrammus sp. (Osteichthyes: Characidae) / Evaluation of formalin and copper sulfate on monogenetic parasite (Urocleidoides sp.) of Hemigrammus sp. (Osteichthyes: Characidae) juveniles

The objective of this study was to evaluate the effectiveness of formalin and copper sulphate to controlm onogeneans injuvenile Hemigrammus spand its effect on leukocyte count. We used acompletely randomized design with five treatments and three replications. All treatments lasted for three days and consisted of: T1, control; T2 and T3 immersion bath for one hour in formalin, an of two concentrations of 0.25ml formalin and the other 0.1mlL-1; the T4 bath for 24 hours with 0,025 ml of formalin-L -1; and T5, bath 24 hours with copper sulphate (0.3 mgL-1). After the experimental period was observed that T2, 100% mortality. The T3 and T5 provided 100% efficiency, however, fish presented mortalities and intoxication of the 66 and 80% respectively. T4 showed an efficacy of 77.7% and decrease in parasitological indices: total number of parasites of 11, 3 ± 9, mean infection intensity of 3.2 ± 2 and a prevalence of 47.1%. As the leukocytes was observed that at T3, T4 and T5 occurred differences in the proportionof lymphocytes, monocytes and neutrophilsas compared to control. The CuSO4 and formalin reduce the parasite load, but presented to be toxicat higher concentrations.

O objetivo deste trabalho foi avaliar a eficácia da formalina e sulfato de cobre no controle de monogenéticos em juvenis de Hemigrammus sp e seu efeito na contagem diferencial de leucócitos. Utilizou-se um delineamento inteiramente casualizado com 5 tratamentos e 3 repetições. Todos os tratamentos tiveram duração de três dias e corresponderam a: T1, controle; T2 e T3 banho por imersão de uma hora em formalina, em duas concentrações uma de 0,25 ml e outra 0,1 ml formalina L-1; o T4, banho de 24 horas com 0,025 ml de formalina L-1; e o T5, banho de 24 horas com sulfato de cobre (0,3 mg L-1). Após o período experimental observou-se que em T2 ocorreu 100% de mortalidade. O T3 e T5 proporcionaram eficácia de 100%, porém os peixes apresentaram intoxicação e mortalidades de 66 e 80%, respectivamente. T4 apresentou eficácia de 77,7% e diminuição dos índices parasitológicos: número total de parasitos de 11,3± 9, intensidade média de infecção de 3,2± 2 e prevalência de 47,1%. Quanto aos leucócitos observou-se que no T3, T4 e T5 ocorreram diferenças na proporção de linfócitos, monócitos e neutrófilos em relação ao controle. O CuSO4 e a formalina reduzem a carga parasitária, mas apresentaram-se tóxicos nas maiores concentrações.

Commonly used metal and crystalline salts in South African traditional medicine.

ETHNOPHARMACOLOGICAL RELEVANCE: Traditional medicines in the form of plants, animals and/or minerals are used by millions of South Africans. There is currently no data regarding the commonly used mineral elements thus the potential benefits or hazards of such products remain unclear. MATERIALS AND METHODS: Metal and crystalline salts were purchased from a rural market (Nongoma, Zululand, South Africa). Information regarding the colloquial name, price and weight was recorded. Energy dispersive X-ray spectroscopy (EDX) was used to quantatively determine the unknown salts. RESULTS: Six widely available salts were analyzed. Ndonya, as it is colloquially known, refers to two products which look identical to the untrained eye-one is dyed table salt and the other is hexavalent chromium. A further product used medicinally, although not widely available, was identified as iron chromite ore. CONCLUSIONS: The array of substances documented, ranging from benign to carcinogenic, stresses the importance of documenting components used in traditional medicine and confirms the necessity to regulate South Africa traditional medicine. Healthcare workers should be aware of the complexities of using such metal salt.

Histopathological effects of waterborne copper nanoparticles and copper sulphate on the organs of rainbow trout (Oncorhynchus mykiss).

It is unclear whether copper nanoparticles are more toxic than traditional forms of dissolved copper. This study aimed to describe the pathologies in gill, gut, liver, kidney, brain and muscle of juvenile rainbow trout, Oncorhynchus mykiss, exposed in triplicate to either a control (no added Cu), 20 or 100 µg l(-1) of either dissolved Cu (as CuSO(4)) or Cu-NPs (mean primary particle size of 87 ± 27 nm) in a semi-static waterborne exposure regime. Fish were sampled at days 0, 4, and 10 for histology. All treatments caused organ injuries, and the kinds of pathologies observed with Cu-NPs were broadly of the same type as CuSO(4) including: hyperplasia, aneurisms, and necrosis in the secondary lamellae of the gills; swelling of goblet cells, necrosis in the mucosa layer and vacuole formation in the gut; hepatitis-like injury and cells with pyknotic nuclei in the liver; damage to the epithelium of some renal tubules and increased Bowman's space in the kidney. In the brain, some mild changes were observed in the nerve cell bodies in the telencephalon, alteration in the thickness of the mesencephalon layers, and enlargement of blood vessel on the ventral surface of the cerebellum. Changes in the proportional area of muscle fibres were observed in skeletal muscle. Overall the data showed that pathology from CuSO(4) and Cu-NPs were of similar types, but there were some material-type effects in the severity or incidence of injuries with Cu-NPs causing more injury in the intestine, liver and brain than the equivalent concentration of CuSO(4) by the end of the experiment, but in the gill and muscle CuSO(4) caused more pathology.

Double tuning a single input probe for heteronuclear NMR spectroscopy at low field.

Applications of PASADENA in biomedicine are continuing to emerge due to recent demonstrations that hyperpolarized metabolic substrates and the corresponding reaction products persist sufficiently long to be detected in vivo. Biomedical applications of PASADENA typically differ from their basic science counterparts in that the polarization endowed by addition of parahydrogen is usually transferred from nascent protons to coupled storage nuclei for subsequent detection on a higher field imaging instrument. These pre-imaging preparations usually take place at low field, but commercial spectrometers capable of heteronuclear pulsed NMR at frequencies in the range of 100 kHz to 1 MHz are scarce though, in comparison to single channel consoles in that field regime. Reported here is a probe circuit that can be used in conjunction with a phase and amplitude modulation scheme we have developed called PANORAMIC (Precession And Nutation for Observing Rotations At Multiple Intervals about the Carrier), that expands a single channel console capability to double or generally multiple resonance with minimal hardware modifications. The demands of this application are geared towards uniform preparation, and since the hyperpolarized molecules are being detected externally at high field, detection sensitivity is secondary to applied field uniformity over a large reaction volume to accommodate heterogeneous chemistry of gas molecules at a liquid interface. The probe circuit was therefore configured with a large (40 mL) Helmholtz sample coil for uniformity, and double-tuned to the Larmor precession frequencies of (13)C/(1)H (128/510 kHz) within a custom solenoidal electromagnet at a static field of 12 mT. Traditional (on-resonant) as well as PANORAMIC NMR signals with signal to noise ratios of approximately 75 have been routinely acquired with this probe and spectrometer setup from 1024 repetitions on the high frequency channel. The proton excitation pulse width was 240 µs at 6.31 W, compared to a carbon-13 pulse width of 220 µs at 2.51 W. When PANORAMIC refocusing waveforms were transmitted at a carrier frequency of 319 kHz, integrated signal intensities from a spin-echo sequence at both proton (510 kHz) and carbon-13 (128 kHz) frequencies were within experimental error to block pulse analogs transmitted on resonance. We anticipate that this probe circuit design could be extended to higher and lower frequencies, and that when used in conjunction with PANORAMIC phase and amplitude modulated arrays, will enable low field imaging consoles to serve as multinuclear consoles.

Effects of copper sulphate concentrations during in vitro maturation of bovine oocytes.

THE OBJECTIVES WERE TO EVALUATE: 1) copper (Cu) concentrations in plasma and follicular fluid (FF) from cattle ovaries; 2) the effects of supplemental Cu during in vitro maturation (IVM) on DNA damage of cumulus cells and glutathione (GSH) content in oocytes and cumulus cells; and 3) supplementary Cu during IVM on subsequent embryo development. Copper concentrations in heifer plasma (116 ± 27.1 µg/dL Cu) were similar (P > 0.05) to concentrations in FF from large (90 ± 20.4 µg/dL Cu) and small (82 ± 22.1 µg/dL Cu) ovarian follicles in these heifers. The DNA damage in cumulus cells decreased with supplemental Cu concentrations of 4 and 6 µg/mL (P < 0.01) in the IVM medium (mean ± SEM index of DNA damage was: 200.0 ± 27.6, 127.6 ± 6.0, 46.4 ± 4.8, and 51.1 ± 6.0 for supplementation with 0, 2, 4, and 6 µg/mL Cu respectively). Total GSH concentrations increased following supplementation with 4 µg/mL Cu (4.7 ± 0.4 pmol in oocytes and 0.4 ± 0.04 nmol/10(6) cumulus cells) and 6 µg/mL Cu (5.0 ± 0.5 pmol in oocytes and 0.5 ± 0.05 nmol/10(6) cumulus cells, P < 0.01) compared with the other classes. Cleavage rates were similar (P ≥ 0.05) when Cu was added to the IVM medium at any concentration (65.1 ± 2.0, 66.6 ± 1.6, 72.0 ± 2.1, and 70.7 ± 2.1 for Cu concentrations of 0, 2, 4, and 6 µg/mL). Percentages of matured oocytes that developed to the blastocyst stage were 18.7 ± 0.6, 26.4 ± 0.03, and 29.0 ± 1.7% for 0, 2, and 4 µg/mL Cu, and was highest (33.2 ± 1.6 %) in oocytes matured with 6 µg/mL Cu (P > 0.01). There was an increase (P > 0.05) in mean cell number per blastocyst obtained from oocytes matured with 4 and 6 µg/mL Cu relative to 0 Cu (IVM alone) and 2 µg/mL Cu. In conclusion, Cu concentrations in the FF and plasma of heifers were similar. Adding copper during oocyte maturation significantly increased both intracellular GSH content and DNA integrity of cumulus cells. Since embryo development was responsive to copper supplementation, we inferred that optimal embryo development to the blastocyst stage was partially dependent on the presence of adequate Cu concentrations during IVM.

Effects of dietary copper supplementation on production performance and plasma biochemical parameters in broiler chickens.

A study was conducted to estimate the effect of copper (Cu) supplementation on growth performance and biochemical profiles of blood and meat in broiler chickens. A total of 240 d-old broiler chicks (Vencobb-100) were randomly divided into 12 groups, each of 20 chicks (4 treatments x 3 replicates). The basal diet (T1) contained 215 g kg⁻¹ crude protein (CP), 12·76 MJ kg⁻¹ ME, 32 g kg⁻¹ total calcium and 5 g kg⁻¹ total phosphorus. T2, T3 and T4 were formulated to contain an additional 75, 150 and 250 mg Cu kg⁻¹ diet, respectively. Copper sulphate pentahydrate (CuSO4, 5H2O) was used as the source of Cu. Significant reductions in plasma total cholesterol and triglyceride, and an elevated concentration of HDL-cholesterol, were observed in the chickens fed with 250 mg Cu kg⁻¹ (T4) of feed at the 3rd and 6th week of the experiment. Total cholesterol in meat decreased significantly in the birds fed with dietary Cu at 250 mg kg⁻¹ (T4) of feed. Growth performance was measured in terms of live weight gain, cumulative feed intake and feed conversion ratio at the end of d 21 and d 42 of the experiment, and the result was found to be commercially beneficial for the chickens receiving 150 mg Cu kg⁻¹ (T3) of diet. The concentration of Cu in breast muscle and liver increased significantly at the end of experiment. From this study it can be concluded that supplementation with dietary Cu may be beneficial for production performance and plasma biochemical characteristics of broiler chickens.