Formulation of Suppositories of Alum Produced from Bauxite Waste in Ghana for the Treatment of Hemorrhoid.

The study sought to formulate and evaluate suppositories using a locally produced brand of alum (Aw) obtained from bauxite waste generated at Awaso bauxite mine in the Western-North region of Ghana, for use in the treatment of hemorrhoids. The suppositories were formulated using shea butter modified, respectively, with amounts of beeswax and theobroma oil. In another development, theobroma oil was modified with different concentrations of beeswax. Drug-base interactions were investigated using attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy. The suppositories were prepared using the hot melt and trituration methods. Quality control checks were carried out on the formulations. The evaluated parameters included physical characteristics (texture, presence or absence of entrapped air, and contraction holes), weight uniformity, disintegration time, drug content, and in vitro release profile of the alum from the formulated suppositories. An in vivo analysis was carried out on the most suitable formulation to ascertain its efficacy on inflamed tissues using croton oil-induced rectal inflammation in a rat model. A critical examination of the ATR-FTIR spectra revealed no drug-base interactions. The suppository formulations passed all Pharmacopoeia stated tests. The in vivo study revealed the use of suppositories ameliorated the croton oil-induced hemorrhoid in the rectoanal region of the rats.

Paeonol alleviates dextran sodium sulfate induced colitis involving Candida albicans-associated dysbiosis.

Inflammatory bowel disease (IBD), which consists of ulcerative colitis (UC) and Crohn's disease (CD), is a chronic inflammatory disorder of the gastrointestinal tract. Occurrence and development of UC have been associated with multiple potential causative factors, which include fungal dysbiosis. Growing evidence reveals that Candida albicans-associated dysbiosis is correlated with clinical deterioration in UC. Paeonol (PAE) is a commonly used traditional medicine with multiple reported properties including effective alleviation of UC. In this study, a murine UC model was established by colonizing mice with additional C. albicans via gavage prior to dextran sodium sulfate (DSS) administration. Effects of PAE treatment were also assessed at initiation and in preestablished C. albicans-associated colitis. The results showed that C. albicans supplementation could aggravate disease activity index (DAI), compromise mucosal integrity, exacerbate fecal and tissue fungal burdens, increase serum ß-glucan and anti-Saccharomyces cerevisiae antibody (ASCA) levels, promote serum and colonic tissue pro-inflammatory cytokine secretion (tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-8) and decrease the anti-inflammatory cytokine IL-10 level. It also stimulated Dectin-1, TLR2 and TLR4 as well as expression of their downstream effector NF-κB in colonic tissue. After PAE treatment, the adverse impacts of C. albicans on colitis were relieved, via decreased receptor-associated local and systemic inflammation. Our study suggests that PAE should be a candidate for treatment of fungal dysbiosis-associated UC and may act through the Dectin-1/NF-κB pathway in collaboration with TLR2 and TLR4. LAY SUMMARY: Candida albicans is believed to be an important stimulator in ulcerative colitice (UC) development. Suppressing the growth of intestinal C. albicans can be contributory to the amelioration of UC. Paeonol (PAE) is a commonly used traditional medicine with multiple biological functions. In this study, we observed that PAE could alleviate symptoms in mice UC model accompanying with burden reduction of C. albicans. Therefore, we suppose that PAE can be a candidate in the treatment of C. albicans-associated UC.

Dietary Supplementation with Sodium Sulfate Improves Rumen Fermentation, Fiber Digestibility, and the Plasma Metabolome through Modulation of Rumen Bacterial Communities in Steers.

Six steers were used to study the effects of dietary supplementation with sodium sulfate (Na2SO4) on rumen fermentation, nutrient digestion, rumen microbiota, and plasma metabolites. The animals were fed a basal ration with Na2SO4 added at 0 g/day (sulfur [S] content of 0.115% dry matter [DM]), 20 g/day (S at 0.185% DM), or 40 g/day (S at 0.255% DM) in a replicate 3-by-3 Latin square design. The results indicated that supplementing with Na2SO4 increased the ruminal concentration of total volatile fatty acids, the molar proportions of acetate and butyrate, the ruminal concentrations of microbial protein, SO42--S, and S2--S, and the digestibility of fiber, while it decreased the molar proportion of propionate and the ruminal concentration of ammonia nitrogen. Supplementing with Na2SO4 increased the diversity and the richness of rumen microbiota and the relative abundances of the phylum Firmicutes and genera Ruminococcus 2, Rikenellaceae RC9 gut group, and Desulfovibrio, whereas it decreased the relative abundances of the phylum Bacteroidetes and genera Prevotella 1, Prevotellaceae UCG-001, and Treponema 2 Supplementing with Na2SO4 also increased the plasma concentrations of amino acids (l-arginine, l-methionine, l-cysteine, and l-lysine), purine derivatives (xanthine and hypoxanthine), vitamins (thiamine and biotin), and lipids (acetylcarnitine and l-carnitine). It was concluded that supplementing the steer ration with Na2SO4 was beneficial for improving the rumen fermentation, fiber digestibility, and nutrient metabolism through modulating the rumen microbial community.IMPORTANCE Essential elements like nitrogen and sulfur greatly affect rumen fermentation and metabolism in ruminants. However, little knowledge is available on the effects of sulfur on the rumen microbiota and plasma metabolome. The results of the present trial demonstrated that supplementing the steer ration with sodium sulfate markedly improved rumen fermentation, fiber digestibility, and metabolism of amino acids, purine derivatives, and vitamins through effects on the ruminal microbiome. The facts obtained from the present trial clarified the possible mechanisms of the positive effects of sulfur on rumen fermentation and nutrient utilization.

Sulfate and hydroxychloride trace minerals in poultry diets - comparative effects on egg production and quality in laying hens, and growth performance and oxidative stress response in broilers.

Two experiments investigated the effect of sulfate and hydroxychloride trace minerals (TM), Zn, Cu, and Mn, in laying hens and broiler chickens. In Expt. 1, Lohmann Brown pullets (total of 1,344) at 21 wk of age were used for a 24-wk experiment. Each of the two treatments had 32 replicates with 21 hens per replicate. At 45 wk of age, three eggs per cage were randomly selected and used for internal quality assessment. In Expt. 2, Ross 308 broilers (total of 1,080) were allocated to two treatments. Each treatment had 30 replicates with 15 chicks per replicate. On day 28, after weighing, three birds were randomly selected from 15 randomly selected pens per treatment. The birds were euthanized and blood was collected for analysis for uric acid, C-reactive protein and methylmalonic acid. Samples were also taken from pectoralis muscle of each chicken and analyzed for mRNA expression of protein synthesis or hydrolysis genes. On day 35, 7 birds per pen were used for carcass evaluation. In Expt. 1, egg weight was greater (P < 0.01) in birds receiving sulfate TM from week 16 (of experiment) onwards whereas the percentage of cracked eggs was lower (P < 0.01) in hens receiving hydroxychloride TM. Percentage hen-day production tended to be greater (P < 0.10) in hens receiving hydroxychloride TM in weeks 4 to 8 only. In Expt. 2, birds receiving hydroxychloride TM had greater (P < 0.05) weight gain and tended to have greater (P < 0.10) feed intake on day 35. Expression of the gene, PSMA1, was lower (P < 0.05) whereas plasma level of uric acid and methyl malonic acid tended to be lower (P < 0.10) in birds receiving hydroxychloride TM. It was concluded that hydroxychloride TM reduced egg loss in hens at peak production and that improved growth performance response in broilers can be partly explained by reduction in proteolytic activities in the pectoralis muscle and greater resilience to oxidative stress.

Effect of manganese preconditioning and replacing inorganic manganese with organic manganese on performance of male broiler chicks.

The effects of manganese (Mn) preconditioning, 96 h post-hatch followed by the replacement of inorganic Mn with different levels of organic Mn (5 to 21 D), on growth, tissue excreta Mn content, gene expression, and enzyme activity were evaluated. A total of 420 day-old male Cobb 500 broilers were divided into 2 groups. One group was fed a corn-soybean meal basal diet containing 17 mg of Mn/kg (preconditioning diet, MnPD); the second group was fed the non-preconditioning diet (NPCD), which was the MnPD supplemented with 60 mg of Mn/kg from manganese sulfate (MnSO4). On day 5, each group was divided into 5 subgroups and were randomly assigned to dietary treatments consisting of MnPD alone or MnPD supplemented with 12 or 60 mg Mn/kg Mn as MnSO4 or Mn proteinate (6 replicate cages of 6 birds). Broiler chicks that were fed the MnPD had lower (P ≤ 0.05) body weight gain (BWG) and G:F ratio when compared to those that were fed the NPCD for 4 D. Birds that were fed MnPD (1 to 4 D) and switched to MnPD supplemented with 60 mg/kg Mn (5 to 21 D) had lower (P ≤ 0.05) BWG compared to those that were fed NPCD (1 to 4 D) and switched to MnPD supplemented with 60 mg/kg Mn for 21 D. Excreta, tibia ash, liver, and heart Mn levels were increased (P ≤ 0.05) by supplemental Mn. The expression of jejunum divalent metal transporter-1 mRNA levels, as well as activities of plasma total super oxide dismutase and liver alanine transaminase, was not affected by MnPD or Mn source and levels. These results confirmed that feeding marginally deficient Mn diets to broiler chicks post-hatch does affect growth rate and tissue Mn concentration.

Dietary manganese supplementation affects mammillary knobs of eggshell ultrastructure in laying hens.

This study evaluated the mechanism by which dietary manganese (Mn) supplementation-in either an organic or inorganic form-affects mammillary knobs of the eggshell ultrastructure in laying hens. A total of 225 54-week-old Hy-Line Brown laying hens were fed a basal diet containing 27.5 mg Mn/kg feed for 2 wk, after which they were randomly allocated into 3 groups and fed a basal diet (control) or a basal diet supplemented with 120 mg Mn/kg feed from monohydrate Mn sulfate (an inorganic source of Mn) or with 80 mg Mn/kg feed from an amino acid-Mn complex (an organic source of Mn) for 10 wk. For each group, 5 replicates of 15 hens each were used with 1 hen per cage. Compared with the control, dietary Mn supplementation increased the mammillary-knob density of eggs at 9.5 h post-oviposition (P < 0.05). The Mn content in both blood and eggshell gland was increased with the supplementation of Mn in inorganic and organic forms (P < 0.05), but the blood Mn content was higher after inorganic-Mn supplementation as compared with organic-Mn supplementation (P < 0.05). RNA sequencing and quantitative real-time PCR analysis of the eggshell gland showed that dietary Mn supplementation increased the expression of genes encoding some proteoglycans, glycoproteins, and calcium-binding proteins in the eggshell gland (P < 0.05), and involved in the process of the protein glycosylation and glycan metabolism in the eggshell gland (P < 0.05). Overall, dietary Mn supplementation can involve in the process of protein glycosylation and glycan metabolism and improve the expression of genes encoding proteoglycans and glycoproteins in the eggshell gland, thus increasing the mammillary-knob density during the initial deposition stage of shell formation.

Interactive effects of supplemental Zn sulfate and ractopamine hydrochloride on growth performance, carcass traits, and plasma urea nitrogen in feedlot heifers.

Interactive effects of supplemental Zn and ractopamine hydrochloride (RH) were evaluated using 156 crossbred heifers (initial BW = 527 kg ± 6.61; gross BW × 0.96) to determine the impact on feedlot performance, plasma urea nitrogen (PUN), and carcass characteristics. The study was conducted as a randomized complete block design with a 2 × 2 factorial arrangement of treatments. Factors consisted of 1) 30 or 100 mg supplemental Zn/kg diet DM (30Zn or 100Zn) as Zn sulfate and 2) 0 or 200 mg RH/heifer daily. Heifers were blocked by BW and assigned randomly within block to treatments for a 43-d trial. Heifers were housed in partially covered feeding pens (3 heifers/pen; 13 pens/treatment) and provided ad libitum access to feed. Ractopamine hydrochloride was fed for 42 d and removed from the diet until cattle were harvested on d 43. Zinc treatments were fed until harvest. Plasma samples were collected on d 0 and 36 to assess changes in plasma Zn and PUN. On d 43, heifers were weighed, then transported to a commercial abattoir where HCW and incidence of liver abscesses were recorded. Carcass data were collected after 32 h of refrigeration. No Zn × RH interactions were observed for plasma Zn or PUN ( ≥ 0.58); however, there was a tendency for a RH × day interaction for PUN ( = 0.08). Supplementing 100Zn resulted in increased plasma Zn ( = 0.02) compared to 30Zn. No RH × Zn interactions were observed for feedlot performance ( ≥ 0.24). Final BW and ADG increased with RH supplementation ( ≤ 0.02), but DMI was not affected ( = 0.63); thus, feed efficiency improved ( < 0.01) when heifers were fed RH. Supplementing 100Zn tended to reduce ADG ( = 0.07) but did not affect other measures of feedlot performance ( ≥ 0.12). Zinc × RH interactions were observed for LM area and yield grade ( ≤ 0.01); LM area decreased and yield grade increased when heifers were supplemented 100Zn with no RH compared to other treatments. A tendency for a Zn × RH interaction was observed for dressed yield ( = 0.08), but no other interactions or effects of Zn were detected for carcass traits ( ≥ 0.11). Supplementing RH increased HCW ( = 0.03) but did not affect other carcass traits ( ≥ 0.13). In conclusion, supplemental Zn had little impact on feedlot performance or PUN concentration but may alter muscle and fat deposition when fed in conjunction with RH.

Effect of feeding Rumen-protected capsule containing niacin, K2SO4, vitamin C, and gamma-aminobutyric acid on heat stress and performance of dairy cows.

This study was conducted to evaluate the effects of supplemental rumen-protected capsule (RPC) on animal performance, serological indicators, and serum heat shock protein 70 (HSP70) of lactating Holstein cows under heat stress (HS). During summer months, 30 healthy multiparous lactating Holstein cows with a parity number of 3.1 ± 0.44, 70 ± 15 d in milk, an average body weight of 622 ± 62kg, and an average milk yield of 32.28 ± 0.96kg/d, were used. The cows were randomly allocated to two groups: a control group and an RPC-supplemented group (0.13373kg K2SO4, 0.02488kg vitamin C, 0.021148kg niacin, and 0.044784kggamma-aminobutyric acid per cow). During the 42-d experiment, ambient air temperature and relative humidity inside and outside the barn were recorded hourly every day for the determination of temperature-humidity index (THI). Milk and blood samples were collected every week, and body weight and body condition scoring were measured on day 0. Based on the THI values, the animals had moderate HS. On day 42, the RPC group had lower HSP70, adrenocorticotropic hormone (P = 0.0001), lactate dehydrogenase (P = 0.0338), and IL-6 (P = 0.0724) levels than the control group, with no significant differences in creatine kinase, glucocorticoid, or IL-2 levels. Milk yield, energy-corrected milk, and dry matter intake were higher in RPC than in the control group (P = 0.0196). There were no significant differences in milk fat or daily protein levels between the two groups; however, daily protein and milk fat levels were higher in the RPC group than in the control group (P = 0.0114 and P = 0.0665, respectively). Somatic cell counts were no different between the two groups. In conclusion, RPC may alleviate HS and improve dairy cow performance.

Effects of hydroxychloride and sulfate form of zinc and manganese supplementation on superoxide dismutase activity and immune responses post lipopolysaccharide challenge in poultry fed marginally lower doses of zinc and manganese.

Effects of hydroxychloride (OHCl) and sulfate form of zinc and manganese supplementation on immune responses of birds fed marginally lower levels of zinc and manganese during an experimental lipopolysaccharide (LPS) injection were studied. In experiment I, 30-week-old layer birds were fed 50 mg/kg Zn+45 mg/kg Mn or 100 mg/kg Zn+90 mg/kg Mn in sulfate or OHCl form and injected with 0 or 500 µg/kg LPS in a 2 (50 mg Zn+45 mg Mn and 100 mg Zn+90 mg Mn) X 2 (sulfate and OHCl) X 2 (0 and 500 µg LPS) factorial setup of treatments for 10 weeks. Among LPS-injected birds, those receiving 50 mg ZnOHCl+45 mg MnOHCl had comparable heterophil and monocyte superoxide dismutase (SOD) activity compared to the birds fed 100 mg Zn+90 mg Mn. Compared to the birds injected with PBS, LPS injection upregulated cathelicidin and IL-1 relative mRNA amounts in monocytes from birds fed 100 mg Zn+90 mg Mn, both in sulfate and OHCl form, and in birds fed 50 mg ZnOHCl+45 mg MnOHCl, but not in the birds fed 50 mg ZnSO4+45 mg MnSO4. In experiment II, one-day-old broiler birds were fed 50 mg ZnOHCl+45 mg MnOHCl, 50 mg ZnOHCl+90 mg MnOHCl, 100 mg ZnOHCL+45 mg MnOHCl, 100 mg ZnOHCl+90 mg MnOHCl, 50 mg ZnSO4+45 mg MnSO4, or 100 mg ZnSO4+90 mg MnSO4 for 21 and 42 days. Birds fed 100 mg ZnOHCl+45 mg MnOHCl form had a comparable heterophil and monocyte SOD activity and monocyte cathelicidin mRNA amounts compared to the group fed 100 mg Zn+90 mg Mn. Increasing the ZnOHCl content from 50 mg to 100 mg/kg Zn reversed (P > 0.05) the decrease in SOD activity and monocyte cathelicidin mRNA levels of the 50 mg ZnOHCl+45 mg MnOHCL fed group, and increasing the MnOHCl content from 45 mg to 90 mg/kg in the 100 mg ZnOHCl+45 mg MnOHCl group further increased SOD activity. In conclusion, birds fed diets with lower amounts of zinc and manganese in sulfate form decreased SOD activity and IL-1 and cathelicidin amounts during inflammation, and either increasing the dietary zinc and manganese content or feeding zinc and manganese in OHCl form synergistically increased the SOD activity and IL-1 and cathelicidin mRNA amounts in immune cells.

Systemic administration of l-kynurenine sulfate induces cerebral hypoperfusion transients in adult C57Bl/6 mice.

The kynurenine pathway is a cascade of enzymatic steps generating biologically active compounds. l-kynurenine (l-KYN) is a central metabolite of tryptophan degradation. In the mammalian brain, l-KYN is partly converted to kynurenic acid (KYNA), which exerts multiple effects on neurotransmission. Recently, l-KYN or one of its derivatives were attributed a direct role in the regulation of the systemic circulation. l-KYN dilates arterial blood vessels during sepsis in rats, while it increases cerebral blood flow (CBF) in awake rabbits. Therefore, we hypothesized that acute elevation of systemic l-KYN concentration may exert potential effects on mean arterial blood pressure (MABP) and on resting CBF in the mouse brain. C57Bl/6 male mice were anesthetized with isoflurane, and MABP was monitored in the femoral artery, while CBF was assessed through the intact parietal bone with the aid of laser speckle contrast imaging. l-KYN sulfate (l-KYNs) (300mg/kg, i.p.) or vehicle was administered intraperitoneally. Subsequently, MABP and CBF were continuously monitored for 2.5h. In the control group, MABP and CBF were stable (69±4mmHg and 100±5%, respectively) throughout the entire data acquisition period. In the l-KYNs-treated group, MABP was similar to that, of control group (73±6mmHg), while hypoperfusion transients of 22±6%, lasting 7±3min occurred in the cerebral cortex over the first 60-120min following drug administration. In conclusion, the systemic high-dose of l-KYNs treatment destabilizes resting CBF by inducing a number of transient hypoperfusion events. This observation indicates the careful consideration of the dose of l-KYN administration by interpreting the effect of kynurenergic manipulation on brain function. By planning clinical trials basing on kynurenergic manipulation possible vascular side effects should also be considered.

Dietary manganese supplementation modulated mechanical and ultrastructural changes during eggshell formation in laying hens.

This study evaluated the mechanical and ultrastructural changes during eggshell formation in laying hens by using the optimal levels of organic and inorganic manganese (Mn). A total of 270 62-wk-old Hy-Line Brown laying hens were fed a basal diet containing 25.1 mg Mn per kg feed for 2 wks, after which they were randomly allocated into 3 groups and fed the basal diet (control) or the basal diet supplemented with 120 mg Mn per kg feed from monohydrate Mn sulfate (an inorganic source of Mn), or 80 mg Mn per kg feed from an amino acid-Mn complex (an organic source of Mn) for 12 wks. For each group, 6 replicates of 15 hens were used with one hen per cage. Dietary Mn supplementation significantly increased eggshell-breaking strength and thickness in laying hens (P < 0.05). In neither was the elasticity of their eggshell membranes, measured during the nucleation and mammillary knob formation stages, affected by dietary Mn compared with the control (P > 0.05), whereas the breaking strength of the eggshells was greater at the linear and terminate deposition stages compared with the control (P < 0.05). Dietary Mn supplementation decreased the width of the mammillary knobs and the proportion of mammillary thickness, and increased the proportion of effective thickness of the whole eggshells (P < 0.05). Ultrastructural changes during the eggshell formation indicated that dietary Mn supplementation increased the nucleation site and mammillary knob densities, decreased the mammillary thickness, and increased the proportion of effective thickness and total thickness of the eggshells compared with the control (P < 0.05). Therefore, dietary Mn supplementation can improve the breaking strength and ultrastructure of the eggshells during their formation, and the mammillary and palisade layers are both crucial structures affected by Mn.

Use of molecular biomarkers to estimate manganese requirements for broiler chickens from 22 to 42 d of age.

The present study was carried out to evaluate dietary Mn requirements of broilers from 22 to 42 d of age using molecular biomarkers. Chickens were fed a conventional basal maize-soyabean meal diet supplemented with Mn as Mn sulphate in graded concentrations of 20 mg Mn/kg from 0 to 140 mg Mn/kg of diet for 21 d (from 22 to 42 d of age). The Mn response curves were fitted for ten parameters including heart Mn-containing superoxide dismutase (MnSOD) mRNA and its protein expression levels and the DNA-binding activities of specificity protein 1 (Sp1) and activating protein-2 (AP-2). Heart MnSOD mRNA and protein expression levels showed significant quadratic responses (P<0·01), and heart MnSOD activity showed a broken-line response (P<0·01), whereas Mn content and DNA-binding activities of Sp1 and AP-2 in the heart displayed linear responses (P<0·01) to dietary Mn concentrations, respectively. The estimates of dietary Mn requirements were 101, 104 and 94 mg/kg for full expressions of MnSOD mRNA level, MnSOD protein level and MnSOD activity in the heart, respectively. Our findings indicate that heart MnSOD mRNA expression level is a more reliable indicator than heart MnSOD protein expression level and its activity for the evaluation of Mn requirement of broilers, and about 100 mg Mn/kg of diet is required for the full expression of heart MnSOD in broilers fed the conventional basal maize-soyabean meal diet from 22 to 42 d of age.

Effect of dietary manganese on antioxidant status and expressions of heat shock proteins and factors in tissues of laying broiler breeders under normal and high environmental temperatures.

To investigate the effect of Mn on antioxidant status and on the expressions of heat shock proteins/factors in tissues of laying broiler breeders subjected to heat challenge, we used a completely randomised design (n 6) with a factorial arrangement of 2 environmental temperatures (normal, 21±1°C, and high, 32±1°C)×3 dietary Mn treatments (a Mn-unsupplemented basal diet (CON), or a basal diet supplemented with 120 mg Mn/kg diet, either as inorganic Mn sulphate (iMn) or as organic Mn proteinate (oMn)). There were no interactions (P>0·10) between environmental temperature and dietary Mn in any of the measured indices. High temperature decreased (P<0·003) Mn content, and also tended (P=0·07) to decrease Cu Zn superoxide dismutase (CuZnSOD) activity in the liver and heart. However, an increased Mn superoxide dismutase (MnSOD) activity (P<0·05) and a slight increase in malondialdehyde level (P=0·06) were detected in breast muscle. Up-regulated (P<0·05) expressions of heat shock factor 1 (HSF1) and HSF3 mRNA and heat shock protein 70 (HSP70) mRNA and protein were found in all three tissues. Broiler breeders fed either iMn or oMn had higher tissue Mn content (P<0·0001), heart MnSOD and CuZnSOD activities (P<0·01) and breast muscle MnSOD protein levels (P<0·05), and lower (P<0·05) breast muscle HSP70 mRNA and protein levels compared with those fed CON. Broiler breeders fed oMn had higher (P<0·03) bone Mn content than those fed iMn. These results indicate that high temperature decreases Mn retention and increases HSP70, HSF1 and HSF3 expressions in the tissues of laying broiler breeders. Furthermore, dietary supplementation with Mn in either source may enhance the heart's antioxidant ability and inhibit the expression of HSP70 in breast muscle. Finally, the organic Mn appears to be more available than inorganic Mn for bone in laying broiler breeders regardless of environmental temperatures.

Dietary manganese source does not affect Mn, Zn and Cu tissue deposition and the activity of manganese-containing enzymes in lambs.

Manganese (Mn) is a trace element required for normal physiological processes in animals and humans. Organic forms of trace elements are expected to have higher bioavailability in comparison with inorganic sources. The effect of feeding a diet supplemented with different sources of manganese to lambs was studied in a 112-d feeding trial. The aim of this study was to investigate the deposition of Mn in relation to activities of superoxide dismutase (SOD), manganese superoxide dismutase (MnSOD) and arginase in the tissues of lambs fed the diet supplemented with an inorganic or an organic source of manganese up to the maximum total Mn content allowed in the European Union (150mg Mn/kg). A total of eighteen female lambs of the improved Valachian breed were randomly allocated to three dietary treatments and fed an unsupplemented basal diet (Control, 31mg Mn/kg) or the identical diet supplemented with manganese sulphate (MnSO4) or manganese chelate of glycine hydrate (Mn-Gly) with a total Mn content up to 150mg/kg. Regardless of the source, feed supplementation with manganese increased Mn concentrations in plasma (P˂0.05) and the liver (P˂0.001) as well as the activity of liver MnSOD (P˂0.05) and arginase (P˂0.001) compared with the control lambs. In the kidney cortex, the concentration of Mn was greatest in lambs fed the diet supplemented with the chelated Mn source compared with animals receiving the inorganic Mn source (P˂0.05) and the unsupplemented diet (P˂0.001). The 112-d intake of feed enriched with manganese did not result in any change in Mn levels, SOD or MnSOD activity in pancreas and kidney tissues. Plasma Cu concentration was depressed in both supplemented treatments. No analyzed tissue showed a change in zinc and copper levels, except the greater Cu concentration in the liver of lambs fed the diet with Mn-Gly. The presented results did not indicate any differences between dietary Mn sources either in Mn tissue deposition or activity of SOD, MnSOD and arginase in lamb tissues; however, it seems that manganese intake of up to 150mg/kg in feed may decrease plasma Cu concentration in lambs.

Macroelemental composition of cadmium stressed lettuce plants grown under conditions of intensive sulphur nutrition.

Lettuce (Lactuca sativa L.) is moderately sensitive to cadmium (Cd) and shows high accumulation of this metal. Thus, this species is considered to be a good model for both identifying determinants controlling Cd accumulation in plant tissues and for developing breeding strategies aimed at limiting the accumulation of this metal in edible tissues. Simultaneously, lettuce is characterised by medium requirements for sulphur (S) - a macronutrient whose role is associated not only with proper growth and development, but also with stress tolerance. The common use of NPK fertilizers without sulphates (S-SO4) together with the progressive process of reducing emissions of S compounds to the natural environment may lead to deficiency of this element in plants. The present study evaluated the changes in macronutrient content and accumulation in Cd-stressed lettuce 'Justyna' supplied with different S doses. Four concentrations of Cd (0, 0.0002, 0.02 or 0.04 mM) and three levels of S applied in the form of S-SO4 (2, 6 or 9 mM S) were used. Cd exposure impaired the macronutrient balance and accumulation in lettuce. Intensive S nutrition to some extent alleviated Cd-induced toxicity. High S doses, especially 6 mM S, partially improved macronutrient status and restored the macronutrient balance. In Cd-stressed plants supplemented with additional S, an increase in root and shoot biomass and in the content of N, K and Mg was found, without significant changes in the Ca content. Simultaneously, the P and S contents in the biomass of both above- and underground organs remained unchanged. In the leaves, as opposite to the roots, intensive S nutrition reduced the accumulation of Cd. However, the foliar Cd concentration still exceeded the acceptable limits established for consumption. All the obtained results concerning the content of macronutrients and their ratios were referred, inter alia, to the standards i.e. the Diagnosis and Recommendation Integrated System (DRIS) norms.

Antitumor activities and immunomodulatory of rice bran polysaccharides and its sulfates in vitro.

Polysaccharides purified from rice bran show antitumor activity against tumor cells, yet the mechanism of this action remains poorly understood. To address this issue, our study evaluated the effect of rice bran polysaccharides on mouse melanoma cell line B16, and Raw264.7 macrophages. Rice bran polysaccharides (RBP) failed to inhibit B16 cell growth in vitro. However, Raw264.7 macrophages treated by RBP enhancement of cytotoxic effects. The cytotoxicity was confirmed by the stimulation of nitric oxide (NO) production and tumor necrosis factor-α (TNF-α) secretion on Raw264.7 macrophages in a dose-dependent manner. RBP2, a fraction of RBP, notably enhanced the inhibition of B16 cells and boosted the immunepotentiation effect compared with RBP. To further enhance the inhibition of B16 cell growth, sulfated polysaccharides (SRBP) was derived using the chlorosulfonic acid-pyridine method. SRBP2 was found to suppress B16 cell growth, reduce B16 cell survival and stimulate NO and TNF-α production. However, SRBP2 displayed a cytotoxic effect on Raw264.7 macrophages. These results suggest that the antitumor activity of RBP and RBP2 is mediated mainly through the activation of macrophages. SRBP2 exerts its antitumor activity by inducing apoptosis in tumor cells and the secretion of NO and TNF-α.

Dietary iron concentration influences serum concentrations of manganese in rats consuming organic or inorganic sources of manganese.

To determine the effects of dietary Fe concentration on Mn bioavailability in rats fed inorganic or organic Mn sources, fifty-four 22-d-old male rats were randomly assigned and fed a basal diet (2·63 mg Fe/kg) supplemented with 0 (low Fe (L-Fe)), 35 (adequate Fe (A-Fe)) or 175 (high Fe (H-Fe)) mg Fe/kg with 10 mg Mn/kg from MnSO4 or Mn-lysine chelate (MnLys). Tissues were harvested after 21 d of feeding. Serum Mn was greater (P<0·05) in MnLys rats than in MnSO4 rats, and in L-Fe rats than in A-Fe or H-Fe rats. Duodenal divalent metal transporter-1 (DMT1) mRNA was lower (P<0·05) in H-Fe rats than in A-Fe rats for the MnSO4 treatment; however, no significant difference was observed between them for MnLys. Liver DMT1 mRNA abundance was greater (P<0·05) in MnSO4 than in the MnLys group for H-Fe rats. The DMT1 protein in duodenum and liver and ferroportin 1 (FPN1) protein in liver was greater (P<0·05) in the MnSO4 group than in the MnLys group, and in L-Fe rats than in H-Fe rats. Duodenal FPN1 protein was greater (P<0·05) in L-Fe rats than in A-Fe rats for the MnLys treatment, but it was not different between them for the MnSO4 treatment. Results suggest that MnLys increased serum Mn concentration as compared with MnSO4 in rats irrespective of dietary Fe concentration, which was not because of the difference in DMT1 and FPN1 expression in the intestine and liver.

Interaction of Formulation and Device Factors Determine the In Vitro Performance of Salbutamol Sulphate Dry Powders for Inhalation.

A variety of capsule-based dry powder inhalers were used to evaluate formulation-device interaction. The in vitro deposition of salbutamol sulphate (SS) was compared directly to published data for salmeterol xinafoate (SX). A 3(2) factorial design was used to assess the effect of SS formulations with three blends of different grade coarse lactose supplemented with different levels of fine lactose. These formulations were tested for homogeneity and evaluated for their in vitro deposition using Aeroliser, Handihaler and Rotahaler devices. The performance of the SS-lactose formulations differed across the grade of lactose and amount of fine lactose used compared to the same powder compositions blended with SX. SX had a greater fine particle fraction than SS for most of the comparable formulations, probably because of the different cohesiveness of the drugs. A head-to-head comparison of 'matched' SX and SS formulations when aerosolised from the same three devices demonstrated that formulation-device interactions are as critical in determining the in vitro deposition of drug-lactose blends as the identity of the active pharmaceutical ingredient. This work has revealed the limitations of the interpretative value of published in vitro performance data generated with a single device (even at equivalent aerosolisation force), when designing formulations for a different device.

Improvement of oxidative and metabolic parameters by cellfood administration in patients affected by neurodegenerative diseases on chelation treatment.

OBJECTIVE: This prospective pilot study aimed at evaluating the effects of therapy with antioxidant compounds (Cellfood, and other antioxidants) on patients affected by neurodegenerative diseases (ND), who displayed toxic metal burden and were subjected to chelation treatment with the chelating agent calcium disodium ethylenediaminetetraacetic acid (CaNa2EDTA or EDTA). METHODS: Two groups of subjects were studied: (a) 39 patients affected by ND and (b) 11 subjects unaffected by ND (controls). The following blood parameters were analyzed before and after three months' treatment with chelation+Cellfood or chelation+other antioxidants: oxidative status (reactive oxygen species, ROS; total antioxidant capacity, TAC; oxidized LDL, oxLDL; glutathione), homocysteine, vitamin B12, and folate. RESULTS: After 3-months' chelation+Cellfood administration oxLDL decreased, ROS levels were significantly lower, and TAC and glutathione levels were significantly higher than after chelation+other antioxidants treatment, both in ND patients and in controls. Moreover, homocysteine metabolism had also improved in both groups. CONCLUSIONS: Chelation+Cellfood treatment was more efficient than chelation+other antioxidants improving oxidative status and homocysteine metabolism significantly in ND patients and controls. Although limited to a small number of cases, this study showed how helpful antioxidant treatment with Cellfood was in improving the subjects' metabolic conditions.