Gluconate-Lactobionate-Dextran Perfusion Solutions Attenuate Ischemic Injury and Improve Function in a Murine Cardiac Transplant Model.

Static cold storage is the cheapest and easiest method and current gold standard to store and preserve donor organs. This study aimed to compare the preservative capacity of gluconate-lactobionate-dextran (Unisol) solutions to histidine-tryptophan-ketoglutarate (HTK) solution. Murine syngeneic heterotopic heart transplantations (Balb/c-Balb/c) were carried out after 18 h of static cold storage. Cardiac grafts were either flushed and stored with Unisol-based solutions with high-(UHK) and low-potassium (ULK) ± glutathione, or HTK. Cardiac grafts were assessed for rebeating and functionality, histomorphologic alterations, and cytokine expression. Unisol-based solutions demonstrated a faster rebeating time (UHK 56 s, UHK + Glut 44 s, ULK 45 s, ULK + Glut 47 s) compared to HTK (119.5 s) along with a better contractility early after reperfusion and at the endpoint on POD 3. Ischemic injury led to a significantly increased leukocyte recruitment, with similar degrees of tissue damage and inflammatory infiltrate in all groups, yet the number of apoptotic cells tended to be lower in ULK compared to HTK. In UHK- and ULK-treated animals, a trend toward decreased expression of proinflammatory markers was seen when compared to HTK. Unisol-based solutions showed an improved preservative capacity compared with the gold standard HTK early after cardiac transplantation. Supplemented glutathione did not further improve tissue-protective properties.

Evaluation of in vitro activity of copper gluconate against SARS-CoV-2 using confocal microscopy-based high content screening.

CONTEXT: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) that emerged late in 2019 is the etiologic agent of coronavirus disease 2019 (Covid-19). There is an urgent need to develop curative and preventive therapeutics to limit the current pandemic and to prevent the re-emergence of Covid-19. This study aimed to assess the in vitro activity of copper gluconate against SARS-CoV-2. METHODS: Vero E6 cells were cultured with or without copper gluconate 18-24 hours before infection. Cells were infected with a recombinant GFP expressing SARS-CoV-2. Cells were infected with a recombinant GFP expressing SARS-CoV-2. Infected cells were incubated in fresh medium containing varying concentration of copper gluconate (supplemented with bovine serum albumin or not) for an additional 48 -h period. The infection level was measured by the confocal microscopy-based high content screening method. The cell viability in presence of copper gluconate was assessed by XTT and propidium iodide assays. RESULTS: The viability of Vero E6 cells exposed to copper gluconate up to 200 µM was found to be similar to that of unexposed cells, but it dropped below 70 % with 400 µM of this agent after 72 h of continuous exposure. The infection rate was 23.8 %, 18.9 %, 20.6 %, 6.9 %, 5.3 % and 5.2 % in cells treated prior infection with 0, 2, 10, 25, 50 and 100 µM of copper gluconate respectively. As compared to untreated cells, the number of infected cells was reduced by 71 %, 77 %, and 78 % with 25, 50, and 100 µM of copper gluconate respectively (p < 0.05). In cells treated only post-infection, the rate of infection dropped by 73 % with 100 µM of copper gluconate (p < 0.05). However, the antiviral activity of copper gluconate was abolished by the addition of bovine serum albumin. CONCLUSION: Copper gluconate was found to mitigate SARS-CoV-2 infection in Vero E6 cells but this effect was abolished by albumin, which suggests that copper will not retain its activity in serum. Furthers studies are needed to investigate whether copper gluconate could be of benefit in mucosal administration such as mouthwash, nasal spray or aerosols.

Effects of zinc supplementation on serum adiponectin concentration and glycemic control in patients with type 2 diabetes.

BACKGROUND: Previous studies have suggested that zinc is involved in insulin homeostasis. Adiponectin is a well-known adipokine with anti-diabetic, anti-atherogenic, and anti-inflammatory properties. The aim of this study was to investigate the effect of zinc supplementation on glycemic control, and the potential mediating role of adiponectin, in patients with type 2 diabetes. METHODS: In this randomized double-blind placebo-controlled clinical trial, 60 patients with diabetes, 30-60 years, were randomized to receive either 30 mg/d zinc (as zinc gluconate) or placebo for 12 weeks. Circulating levels of adiponectin, zinc, glucose homeostasis parameters, and lipid profiles, as well as anthropometric parameters and dietary intakes, were assessed. RESULTS: About 53.3% of the patients had zinc insufficiency at baseline. Serum zinc levels improved significantly in the intervention than control group following 12 weeks supplementation (P < 0.001). Adiponectin (1.23 ± 2.23 µg/ml, P = 0.006) and insulin (3.6 ± 4.66 µIU/ml, P = 0.001) levels increased significantly compared to baseline in the zinc group; but this change was not significant compared with the control group. Following supplementation, there were no significant differences in glycemic control and anthropometric parameters between the two groups. Serum HDL levels increased significantly in the zinc (5.37 ± 14.8 mg/dl) compared to control (-1.53 ± 6.9 mg/dl) group following supplementation (P = 0.039). CONCLUSION: Despite a significant increase in serum zinc level, no improvement was observed in glycemic control, following 12 weeks supplementation with 30 mg/d zinc (as zinc gluconate). Zinc supplementation restored adiponectin concentrations partly within the intervention group, and increased HDL levels compared to the control group. The current findings did not support improvement in glucose homeostasis following zinc supplementation in patients with type 2 diabetes under the present study design.

Effects of oral selenium and magnesium co-supplementation on lipid metabolism, antioxidative status, histopathological lesions, and related gene expression in rats fed a high-fat diet.

BACKGROUND: Supplementation with Selenium (Se) has been shown to lower blood cholesterol and increase tissue concentrations of the antioxidant glutathione (GSH); however, the effects of Se supplementation, in combination with supplemental magnesium, on high fat-induced hyperlipidemia have not been studied. This study was designed to elucidate the effects of oral selenium and magnesium co-supplementation on antihyperlipidemic and hepatoprotective, antioxidative activities, and related gene expression in a hyperlipidemic rat model. METHODS: Forty male Sprague Dawley rats were divided into 4 groups: one group served as control group (CT), provided control diet; The other groups were made hyperlipidemic with high-fat diet; specifically, a high-fat diet group (HF); low-dose selenium (0.05 mg/kg·bw) + low-dose magnesium (5.83 mg/kg·bw) supplement high-fat diet group (HF + LSe + LMg) and high-dose selenium (0.10 mg/kg·bw) + high-dose magnesium (58.33 mg/kg·bw) supplement high-fat diet group (HF + HSe + HMg). The first 4 weeks of the experiment was a hyperlipidemia inducing period using high-fat diet and the following 8 weeks involved in selenium and magnesium co-supplementation. On day 0, 20, 40 and 60 of the intervention, lipid profile was measured. At the end of the 12-week experiments, final blood and liver samples were collected for the measurements of lipid profile, antioxidative indexes, pathological examination, and liver lipid metabolism related gene expression. RESULTS: The elevated levels of serum and liver total cholesterol (TC) and serum LDL-C induced by feeding high-fat diets were significantly reduced by low-dose Se and Mg co-supplementation. Both doses of selenium and magnesium co-supplementation notably decreased the blood and liver TG levels, liver function indexes ALT and AST and the ratio of TC/HDL-C and TG/HDL-C. In contrast, Se and Mg supplementation showed a substantial increase in Se-dependent glutathione peroxidase (GSH-Px) and SOD activities and an significant reduce of level of MDA of hyperlipidemic rats. Oil Red O staining showed that selenium and magnesium co-supplementation significantly reduced hepatic intracellular triacylglycerol accumulation. H&E staining also showed that selenium and magnesium co-supplementation can attenuate liver steatosis. Selenium and magnesium co-supplementation remarkably inhibited the mRNA expression level of hepatic lipogenesis genes liver X receptor alpha (LXRα),SREBP-1c and FASN (fatty acid synthase), regulated the mRNA expression levels of liver enzymes related to cholesterol metabolism, including the down regulation of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) and the upregulation of cholesterol 7α-hydroxylase (CYP7A1) and lecithin cholesterol acyltransferase (LCAT) in the liver of hyperlipidemia rats. CONCLUSIONS: Oral selenium and magnesium co-supplementation inhibited an increase of lipid and liver profile and liver function index induced by a high-fat diet, and enhanced the activity of the antioxidant enzymes. Selenium combined with magnesium is a promising therapeutic strategy with lipid-lowering and antioxidative effects that protects the liver against hyperlipidemia.

Metabolic effect of alkaline additives and guanosine/gluconate in storage solutions for red blood cells.

BACKGROUND: Over a century of advancements in the field of additive solutions for red blood cell (RBC) storage has made transfusion therapy a safe and effective practice for millions of recipients worldwide. Still, storage in the blood bank results in the progressive accumulation of metabolic alterations, a phenomenon that is mitigated by storage in novel storage additives, such as alkaline additive solutions. While novel alkaline additive formulations have been proposed, no metabolomics characterization has been performed to date. STUDY DESIGN AND METHODS: We performed UHPLC-MS metabolomics analyses of red blood cells stored in SAGM (standard additive in Europe), (PAGGSM), or alkaline additives SOLX, E-SOL 5 and PAG3M for either 1, 21, 35 (end of shelf-life in the Netherlands), or 56 days. RESULTS: Alkaline additives (especially PAG3M) better preserved 2,3-diphosphoglycerate and adenosine triphosphate (ATP). Deaminated purines such as hypoxanthine were predictive of hemolysis and morphological alterations. Guanosine supplementation in PAGGSM and PAG3M fueled ATP generation by feeding into the nonoxidative pentose phosphate pathway via phosphoribolysis. Decreased urate to hypoxanthine ratios were observed in alkaline additives, suggestive of decreased generation of urate and hydrogen peroxide. Despite the many benefits observed in purine and redox metabolism, alkaline additives did not prevent accumulation of free fatty acids and oxidized byproducts, opening a window for future alkaline formulations including (lipophilic) antioxidants. CONCLUSION: Alkalinization via different strategies (replacement of chloride anions with either high bicarbonate, high citrate/phosphate, or membrane impermeant gluconate) results in different metabolic outcomes, which are superior to current canonical additives in all cases.

Effect of magnesium gluconate administration on lipid metabolism, antioxidative status, and related gene expression in rats fed a high-fat diet.

To explore the effect of magnesium gluconate (MgG) on lipid metabolism and its regulation mechanism through animal experiments, and to provide basis for MgG dietary intervention in hyperlipidemia. The first four weeks was hyperlipidemia-inducing period through high-fat diet and the following eight weeks was the MgG supplementation. At the end of the experiment, blood and liver samples were collected for the measurements of lipid profile, antioxidative indexes, pathological examination, and cholesterol metabolism-related gene expression. Oral administration of MgG notably decreased the blood levels of TC, TG, LDL-C and liver function index ALT and AST of hyperlipidemic rats. The rats supplemented with magnesium showed a huge increase in the GSH-Px and SOD activities, and reduced the heart weight and liver lipid accumulation of high-fat diet fed rats. MgG remarkably up-regulated the mRNA expression levels of LDLR and CYP7A1 of liver enzymes related to cholesterol metabolism. Oral magnesium supplementation inhibited an increase in lipid profile and liver function index by a high-fat diet, and enhanced the activity of the antioxidant enzymes. Magnesium has lipid-lowering and antioxidative effects that protect the liver against hyperlipidemia.

Creatine salts provide neuroprotection even after partial impairment of the creatine transporter.

Creatine, a compound that is critical for energy metabolism of nervous cells, crosses the blood-brain barrier (BBB) and the neuronal plasma membrane with difficulty, and only using its specific transporter. In the hereditary condition where the creatine transporter is defective (creatine transporter deficiency) there is no creatine in the brain, and administration of creatine is useless lacking the transporter. The disease is severe and incurable. Creatine-derived molecules that could cross BBB and plasma membrane independently of the transporter might be useful to cure this condition. Moreover, such molecules could be useful also in stroke and other brain ischemic conditions. In this paper, we investigated three creatine salts, creatine ascorbate, creatine gluconate and creatine glucose. Of these, creatine glucose was ineffective after transporter block with guanidine acetic acid (GPA) administration. Creatine ascorbate was not superior to creatine in increasing tissue creatine and phosphocreatine content after transporter impairment, however even after such impairment it delayed synaptic failure during anoxia. Finally, creatine gluconate was superior to creatine in increasing tissue content of creatine after transporter block and slowed down PS disappearance during anoxia, an effect that creatine did not have. These findings suggest that coupling creatine to molecules having a specific transporter may be a useful strategy in creatine transporter deficiency. In particular, creatine ascorbate has effects comparable to those of creatine in normal conditions, while being superior to it under conditions of missing or impaired creatine transporter.

Effects of surgical and chemical sterilization on the behavior of free-roaming male dogs in Puerto Natales, Chile.

Population management of free-roaming domestic dogs (Canis lupus familiaris) is of interest due to the threat these animals pose to people, other animals and the environment. Current sterilization procedures for male dogs include surgical and chemical methods. However, little is known about how these procedures affect their behavior. The primary objective of this study was to investigate changes in selected behaviors following chemical and surgical sterilization in a male free-roaming dog (FRD) population in southern Chile. We also examined the association between serum testosterone levels and behaviors thought to be influenced by circulating androgens. A total of 174 dogs were randomly assigned to either a surgical or chemical sterilization group, or a control group. At the onset of the intervention period, 119 dogs remained and 102 dogs successfully completed the study. Each dog was monitored pre- and post-intervention using video recordings, GPS collars, and blood samples for the measurement of testosterone. Analysis of behavior revealed that surgically castrated dogs showed no reduction of sexual activity or aggression when compared to their pre-intervention behavior. Chemically sterilized dogs showed a statistically significant increase in dog-directed aggression, but no change in sexual activity. There was no change in home range size in any groups between the pre- and post-intervention measurement. We found no consistent association between levels of serum testosterone concentration and behavioral changes in any of the groups. This study presents the first detailed behavioral observations following surgical and chemical sterilization in male FRDs. The information generated is highly relevant to communities struggling with the control of FRDs. Complementary studies to further our understanding of the effects of male sterilization on the behavioral and reproductive dynamics of FRD populations are needed.

Possible Protective Effects of Quercetin and Sodium Gluconate Against Colon Cancer Induction by Dimethylhydrazine in Mice.

Micronutrients in food have been found to have chemopreventive effects, supporting the conclusions from epidemiologie studies that consumption of fresh fruits and vegetables reduces cancer risk. The present study was carried out to evaluate the role of querctin (Q) and sodium gluconate (GNA) supplementation separately or in combination in ameliorating promotion of colon tumor development by dimethyl-hydrazine (DMH) in mice. Histopathological observation of colons in mice treated with DMH showed goblet cell dysplasia with inflammatory cell infiltration. This pathological finding was associated with significant alteration in oxidative stress markers in colon tissues and carcinoembryonic antigen (CEA) levels in plasma. Mice co-treated with GNA and Q showed mild changes of absorptive and goblet cells and inflammatory cell infiltration in lamina properia, with improvement in oxidative stress markers. In conclusion, findings of the present study indicate significant roles for reactive oxygen species (ROS) in pathogenesis of DMH-induced colon toxicity and initiation of colon cancer. Also, they suggest that Q, GNA or the combination of both have a positive beneficial effect against DMH induced colonic cancer induction in mice.

Moderately High Dose Zinc Gluconate or Zinc Glycinate: Effects on Plasma Zinc and Erythrocyte Superoxide Dismutase Activities in Young Adult Women.

Some zinc (Zn) research studies have used either Zn gluconate or Zn glycinate, but the two forms have not been compared much. Therefore, a moderately high dose of the two forms (60 mg Zn/day) were compared in a 6-week intervention in young adult women. Plasma Zn, the traditional assessment of Zn status, was increased in all subjects given Zn glycinate (N = 10), while no significant change was seen overall for Zn gluconate or placebo (N = 10 each). Erythrocyte superoxide dismutase activity, a marker for Zn-induced copper deficiency, was unchanged in all three groups. Thus, for the conditions of this study, Zn glycinate effectively changed Zn status better than Zn gluconate, but neither impacted copper status.

GPI/AMF inhibition blocks the development of the metastatic phenotype of mature multi-cellular tumor spheroids.

Epithelial-mesenchymal transition (EMT) and cellular invasiveness are two pivotal processes for the development of metastatic tumor phenotypes. The metastatic profile of non-metastatic MCF-7 cells growing as multi-cellular tumor microspheroids (MCTSs) was analyzed by determining the contents of the EMT, invasive and migratory proteins, as well as their migration and invasiveness potential and capacity to secrete active cytokines such as the glucose phosphate isomerase/AMF (GPI/AMF). As for the control, the same analysis was also performed in MCF-7 and MDA-MB-231 (highly metastatic, MDA) monolayer cells, and in stage IIIB and IV human metastatic breast biopsies. The proliferative cell layers (PRL) of mature MCF-7 MCTSs, MDA monolayer cells and metastatic biopsies exhibited increased cellular contents (2-15 times) of EMT (ß-catenin, SNAIL), migratory (vimentin, cytokeratin, and fibronectin) and invasive (MMP-1, VEGF) proteins versus MCF-7 monolayer cells, quiescent cell layers of mature MCF-7 MCTS and non-metastatic breast biopsies. The increase in metastatic proteins correlated with substantially elevated cellular abilities for migration (18-times) and invasiveness (13-times) and with the higher level (6-times) of the cytokine GPI/AMF in the extracellular medium of PRL, as compared to MCF-7 monolayer cells. Interestingly, the addition of the GPI/AMF inhibitors erythrose-4-phosphate or 6-phosphogluconate at micromolar doses significantly decreased its extracellular activity (>80%), with a concomitant diminution in the metastatic protein content and migratory tumor cell capacity, and with no inhibitory effect on tumor lactate production or toxicity on 3T3 mouse fibroblasts. The present findings provide new insights into the discovery of metabolic inhibitors to be used as complementary therapy against metastatic and aggressive tumors.

Effect of zinc supplementation on superoxide dismutase activity in patients with ulcerative rectocolitis.

INTRODUCTION: Ulcerative rectocolitis is characterized by diffuse mucosal inflammation and oxidative stress. Thus, the organism activates the antioxidant defence system in an attempt to reduce the excessive production of reactive oxygen species or neutralize them. OBJECTIVE: This study evaluated the effect of zinc supplementation on the activity of the enzyme superoxide dismutase (SOD) in patients with ulcerative rectocolitis. METHODS: The study included 24 patients, aged between 20 and 59 years and diagnosed with ulcerative rectocolitis, in the remission stage of the disease, who were divided into two groups: experimental - deficient in zinc (n=12) and control - normal or high zinc (n=12). Only the first group underwent supplement intervention, in the form of zinc gluconate (30 mg Zn/day), taken daily in the morning, fasted for 60 days. Plasma and erythrocyte zinc concentrations were determined by flame atomic absorption spectrophotometer. The erythrocyte SOD activity was determined in vitro according to the methodology recommended by the manufacturer Randox. RESULTS AND DISCUSSION: Zinc supplementation caused a significant increase in the plasma concentrations of the mineral, and showed a significant reduction in erythrocyte zinc, remaining within normal limits. The SOD activity was high in patients of both the experimental and control groups, with no difference after supplementation. CONCLUSION: This study demonstrates that zinc supplementation improves the homeostatic condition of the mineral, with no change in SOD activity, as a marker of oxidative stress in patients with ulcerative rectocolitis.

Introducción: La colitis ulcerosa se caracteriza por la inflamación difusa de la mucosa y el estrés oxidativo. De esta forma, el cuerpo activa el sistema de defensa antioxidante en un intento de reducir la producción excesiva de especies reactivas de oxígeno, así como poder neutralizarlos. Objetivo: Este estudio evaluó el efecto de la suplementación de zinc sobre la actividad de la enzima superóxido dismutasa en pacientes con colitis ulcerosa. Métodos: El estudio incluyó 24 pacientes, con edades comprendidas entre 20 y 59 años y con diagnóstico de colitis ulcerosa en fase de remisión de la enfermedad. Los pacientes fueron divididos en dos grupos: experimental - deficiencia de zinc (n = 12) y control - normales o con altos contenido de zinc (n = 12). El grupo experimental se sometió a tratamiento con suplemento de drogas, en forma de gluconato de zinc (30 mg Zn / día), administrada diariamente por la mañana en ayunas durante 60 días. Las concentraciones en plasma y los eritrocitos de zinc se determinaron por espectrofotometría de absorción atómica de llama. La actividad de la superóxido dismutasa (SOD) se determinó por el método de eritrocitos in vitro utilizando el kit de Randox. Resultados y Discusión: La suplementación de zinc causó un aumento significativo en las concentraciones plasmáticas de mineral y mostró una reducción significativa en los eritrocitos, permaneciendo dentro de los límites normales. La actividad de SOD fue mayor en los pacientes de los grupos experimentales y de control, sin diferencias después de la suplementación. Conclusión: El estudio evidenció que la administración de suplementos de zinc mejora la condición homeostática del mineral, sin ningún cambio en la actividad de SOD, como un marcador de estrés oxidativo en pacientes con colitis ulcerosa.

Lack of protective effects of zinc gluconate against rat colon carcinogenesis.

Zinc has been proposed as a promising chemopreventive candidate against colon cancer. However, few studies on the potential beneficial effects of this trace element on cancer chemoprevention are available. The present study was designed to investigate the potential modifying influence of zinc gluconate (ZnGly) on the initiation step of colon carcinogenesis induced by 1,2-dimethylhydrazine (DMH). Male Wistar rats received orally ZnGly (15 mg elemental zinc/kg, 3 times per wk) 2 wk before and during DMH treatment (3 × 40 mg/kg, once a wk). The animals were euthanized at the end of 4th and 16th wk. Colons were analyzed for aberrant crypt foci (ACF) and tumor development. Blood and colon zinc levels, cell proliferation, and apoptosis indexes in colonic crypts were analyzed 24 h after the last DMH administration. Oral treatment with ZnGly did neither alter the number of ACF nor the indexes of cell proliferation and apoptosis in the colonic mucosa. The incidence and multiplicity of colon tumors induced by DMH and their histopathological patterns were not modified by previous treatment with ZnGly. These findings indicate a lack of chemopreventive action of zinc gluconate supplementation on the initiation step of rat colon carcinogenesis induced by DMH.

Significant improvement in islet yield and survival with modified ET-Kyoto solution: ET-Kyoto/Neutrophil elastase inhibitor.

Although islet transplantation can achieve insulin independence in patients with type 1 diabetes, sufficient number of islets derived from two or more donors is usually required to achieve normoglycemia. Activated neutrophils and neutrophil elastase (NE), which is released from these neutrophils, can directly cause injury in islet grafts. We hypothesized that inhibition of NE improves islet isolation and islet allograft survival. We tested our hypothesis by examining the effects of modified ET-Kyoto solution supplemented with sivelestat, a NE inhibitor (S-Kyoto solution), on islet yield and viability in islet isolation and the effect of intraperitoneally injected sivelestat on islet graft survival in a mouse allotransplant model. NE and proinflammatory cytokines such as tumor necrosis factor (TNF)-α and interleukin (IL)-6 increased markedly at the end of warm digestion during islet isolation and exhibited direct cytotoxic activity against the islets causing their apoptosis. The use of S-Kyoto solution significantly improved islet yield and viability. Furthermore, treatment with sivelestat resulted in significant prolongation of islet allograft survival in recipient mice. Furthermore, serum levels of IL-6 and TNF-α at 1 and 2 weeks posttransplantation were significantly higher in islet recipients than before transplantation. Our results indicated that NE released from activated neutrophils negatively affects islet survival and that its suppression both in vitro and in vivo improved islet yield and prolonged islet graft survival. The results suggest that inhibition of NE activity could be potentially useful in islet transplantation for patients with type 1 diabetes mellitus.

Effect of dietary zinc and phytase supplementation on botulinum toxin treatments.

PURPOSE: To determine whether oral zinc supplementation might affect the efficacy and duration of botulinum toxin treatments. METHODS: In a double-blind, placebo-controlled, crossover pilot study, we examined the efficacy of three botulinum toxin preparations (onabotulinumtoxinA, abobotulinumtoxinA, and rimabotulinumtoxinB) following oral supplementation with zinc citrate 50 mg and phytase 3,000 PU, zinc gluconate 10 mg, or lactulose placebo in individuals treated for cosmetic facial rhytids, benign essential blepharospasm, and hemifacial spasm. RESULTS: In seventy-seven patients, 92% of subjects supplemented with zinc 50 mg and phytase experienced an average increase in toxin effect duration of nearly 30%, and 84% of participants reported a subjective increase in toxin effect, whereas no significant increase in duration or effect was reported by patients following supplementation with lactulose placebo or 10 mg of zinc gluconate. The dramatic impact of the zinc/phytase supplementation on some patients' lives clinically unmasked the study and prompted an early termination. CONCLUSIONS: This study suggests a potentially meaningful role for zinc and/or phytase supplementation in increasing the degree and duration of botulinum toxin effect in the treatment of cosmetic facial rhytids, benign essential blepharospasm, and hemifacial spasm.

A comparative study of antioxidative activity of calcium-D-glucarate, sodium-D-gluconate and D-glucono-1,4-lactone in a human blood platelet model.

D-glucono-1,4-lactone, sodium D-gluconate and calcium D-glucarate are non-toxic glucose derivatives occurring naturally in fruits and vegetables. Calcium D-glucarate is promoted as an orally bioavailability dietary supplement with potential chemopreventive activity without adverse effects. Despite many commercial applications in pharmaceutical and food industries the potential activity mechanisms of glucarate and gluconate are not clear. The purpose of this study was to investigate and compare the effects of these compounds on blood platelets under oxidative stress conditions and to examine their role in thrombin-induced platelet activation. Platelet activation is essential in haemostasis, tumor progression and allergic and non-allergic inflammation, where reactive oxygen species are involved. The antiplatelet and antioxidative activity was studied in vitro by measuring levels of specific oxidative stress markers: thiobarbituric acid reactive substances, superoxide anion, carbonyl groups, 3-nitrotyrosine, protein and low molecular weight thiols. All tested compounds significantly inhibited thrombin-induced arachidonic peroxidation, O2⁻ⁱ production and also platelet protein oxidation/nitration induced by peroxynitrite, which is a strong oxidant formed intravascularly in vivo. Carbonyl group generation, thiol oxidation and nitrotyrosine formation were significantly decreased in the presence of glucose derivatives. The obtained results demonstrate that tested compounds may be helpful in the prevention of excessive platelet activation through the antioxidant mechanisms. Comparative studies indicate the predominant preventive activity of sodium D-gluconate. In general, the consumption of apples or apple juice as well as oranges, grapefruit and cruciferous vegetables, sources of large amounts of tested derivatives, have beneficial effects on platelets under oxidative stress.

Influence of chemical activation of a 35% hydrogen peroxide bleaching gel on its penetration and efficacy--in vitro study.

OBJECTIVES: The aim of this study was to evaluate the effects of chemical activation of hydrogen peroxide (HP) gel on colour changes and penetration through the tooth structure. METHODS: One hundred and four bovine incisors were used. One dentine (CD) disc and one enamel-dentine (ED) disc were prepared from each tooth. They were positioned over artificial pulpal chambers and the bleaching was performed with an experimental 35% HP gel. Two control and six experimental groups were prepared. In the positive control group (PC) no chemical activator was used. In the negative control group (NC) the specimens did not receive any bleaching. Each experimental group received a different chemical activator (manganese gluconate-MG; manganese chlorite-MC; ferrous sulphate-FS; ferrous chlorite-FC; and mulberries root extract-MRE). After the bleaching procedure a sample of solution was collected from the artificial pulpal chamber and the HP concentration was measured. The data were analysed using ANOVA, Tukey's, and Dunnett's tests. RESULTS: The groups MG and FS showed a significantly lower penetration of HP than the PC group. For the parameter Delta E, all the groups, with the exception of the group MRE, showed a significantly higher means in relation to the PC group in ED colour. For dentine colour, just the groups MG and FS had significant differences in relation to PC. CONCLUSIONS: The addition of MG and FS decreases the penetration of HP. The chemical activation using metal salts tested was effective in increasing the bleaching effect.

In vitro antioxidant and in vivo prophylactic effects of two gamma-lactones isolated from Grewia tiliaefolia against hepatotoxicity in carbon tetrachloride intoxicated rats.

Grewia tiliaefolia is widely used in traditional Indian medicines to cure jaundice, biliousness, dysentery and the diseases of blood. Bioassay-guided fractionation of methanolic extract of the G. tiliaefolia bark has resulted in the isolation of D-erythro-2-hexenoic acid gamma-lactone (EHGL) and gulonic acid gamma-lactone (GAGL). Hepatoprotective activity of the methanolic extract and the isolated constituents were evaluated against CCl(4)-induced hepatotoxicity in rats. The treatment with methanolic extract, EHGL and GAGL at oral doses of 100, 150 and 60 mg/kg respectively with concomitant CCl(4) intraperitoneal injection (1 ml/kg) significantly reduced the elevated plasma levels of aminotransferases, alkaline phosphatase and the incidence of liver necrosis compared with the CCl(4)-injected group without affecting the concentrations of serum bilirubin and hepatic markers. EHGL and GAGL significantly inhibited the elevated levels of thiobarbituric acid reactive substances and glutathione in liver homogenates. Histology of the liver tissues of the extract and isolated constituents treated groups showed the presence of normal hepatic cords, absence of necrosis and fatty infiltration as similar to the normal control. The results revealed that the hepatoprotective activity of EHGL is significant as similar to the standard drug silymarin. To clarify the influence of the extract and isolated constituents on the protection of oxidative-hepatic damage, we examined in vitro antioxidant properties of the test compounds. The extract and the constituents showed significant free radical scavenging activity. These results suggest that the extract as well as the constituents could protect the hepatocytes from CCl(4)-induced liver damage perhaps, by their anti-oxidative effect on hepatocytes, hence eliminating the deleterious effects of toxic metabolites from CCl(4).

Effect of prenatal zinc supplementation on birthweight.

Although iron and zinc deficiencies are known to occur together and also appear to be high in Ghana, a few supplementation studies addressed this concurrently in pregnancy. In a double-blind, randomized controlled trial, 600 pregnant women in Ghana were randomly assigned to receive either a combined supplement of 40 mg of zinc as zinc gluconate and 40 mg of iron as ferrous sulphate or 40 mg of elemental iron as ferrous sulphate. Overall, there was no detectable difference in the mean birthweight between the study groups, although the effect of iron-zinc supplementation on the mean birthweight was masked by a strong interaction between the type of supplement and the iron status of participants [F (1,179) = 5.614, p = 0.019]. Prenatal iron-zinc supplementation was effective in increasing the mean birthweight among anaemic and iron-deficient women but not among women with elevated iron stores in early pregnancy.

The effect of Grobiotic-P combined with yeast cell wall and gluconic acid on growth performance, nutrient digestibilities, and cecal microbial populations in young chicks.

Two experiments were conducted with New Hampshire x Columbian chicks fed a corn-soybean meal diet to examine the efficacy of varying levels and combinations of Grobiotic P (GB), a prebiotic-type product that contains dairy and yeast fractions and dried fermentation extracts, gluconic acid, and yeast cell wall (YCW) on growth performance, nutrient digestibility, and cecal microbial populations. In experiment 1, chicks were allowed ad libitum access to a corn-soybean meal basal diet or the basal diet supplemented with 2 or 4% GB, 1.5 or 3% gluconic acid, 0.2% YCW, or various 2-way combinations of the supplements in place of dextrose and arenaceous flour. In experiment 2, the same supplement combinations were used as in experiment 1; however, the 3% gluconic acid supplement was eliminated. In both experiments, supplementing GB, YCW, or 1.5% gluconic acid to the diet had no consistent effect on growth performance except that the weight gain was depressed (P < 0.05) when the chicks were fed a diet containing 3% gluconic acid. Chicks fed diets containing 2 or 4% GB combinations showed a significant increase (P < 0.05) in ME(n) at 21 d when compared with chicks fed the basal diet. For amino acid (AA) digestibility, 2% GB combined with 0.2% YCW or 1.5% gluconic acid resulted in a reduction (P < 0.05) in AA digestibility at 7 d; however, at 21 d, these combinations increased (P < 0.05) digestibility of all AA. There was no consistent effect of dietary treatment on cecal bifidobacteria, lactobacilli, Escherichia coli, or Clostridium perfringens populations or cecal pH. These experiments indicated that diets containing GB generally increased ME(n) and AA digestibility at 21 d and that the prebiotic combinations had no consistent effects on cecal microbial populations.