Tumor Microenvironment-Responsive Nanococktails for Synergistic Enhancement of Cancer Treatment via Cascade Reactions.

A combination treatment strategy that relies on the synergetic effects of different therapeutic approaches has been considered to be an effective method for cancer therapy. Herein, a chemotherapeutic drug (doxorubicin, Dox) and a manganese ion (Mn2+) were co-loaded into regenerated silk fibroin-based nanoparticles (NPs), followed by the surface conjugation of phycocyanin (PC) to construct tumor microenvironment-activated nanococktails. The resultant PC-Mn@Dox-NPs showed increased drug release rates by responding to various stimulating factors (acidic pH, hydrogen peroxide (H2O2), and glutathione), revealing that they could efficiently release the payloads (Dox and Mn2+) in tumor cells. The released Dox could not only inhibit the growth of tumor cells but also generated a large amount of H2O2. The elevated H2O2 was decomposed into the highly harmful hydroxyl radicals and oxygen through an Mn2+-mediated Fenton-like reaction. Furthermore, the generated oxygen participated in photodynamic therapy (PDT) and produced abundant singlet oxygen. Our investigations demonstrate that these PC-Mn@Dox-NPs exhibit multiple bioresponsibilities and favorable biosafety. By integrating Dox-induced chemotherapy, Mn2+-mediated chemodynamic therapy, and PC-based PDT via cascade reactions, PC-Mn@Dox-NPs achieved enhanced in vitro and in vivo anticancer efficacies compared to all the mono- or dual-therapeutic approaches. These findings reveal that PC-Mn@Dox-NPs can be exploited as a promising nanococktail for cascade reaction-mediated synergistic cancer treatment.

Metabolomic investigation of regional brain tissue dysfunctions induced by global cerebral ischemia.

BACKGROUND: To get a broader view of global ischemia-induced cerebral disorders at the metabolic level, a nuclear magnetic resonance-based metabolomic study was performed to evaluate the metabolic profile changes on regional brain tissues of female and male mice upon bilateral common carotid arteries occlusion (BCCAO) operation. RESULTS: Significant metabolic disorders were observed in both cerebral cortex and hippocampus tissues of the experimental mice upon global cerebral ischemic attack. Multiple amino acids were identified as the dominantly perturbed metabolites. It was also shown that although the metabolic profile change patterns in the brain tissues were quite similar in male and female BCCAO mice, metabolic disorders in the cortex tissues were more severe in the female mice than in the male mice. CONCLUSIONS: In the present study, significant changes in amino acid metabolic pathways were confirmed in the early stage of global ischemia. Meanwhile, cerebral metabolic dysfunctions were more severe in the female BCCAO mice than in the male mice, suggesting that gender may play a role in different metabolic responses to the ischemic attack, which may provide an important hypothesis for a better understanding of the clinically observed gender-dependent pathological outcome of cerebral ischemia.

Application of Polymeric Nanoparticles for CNS Targeted Zinc Delivery In Vivo.

A dyshomeostasis of zinc ions has been reported for many psychiatric and neurodegenerative disorders including schizophrenia, attention deficit hyperactivity disorder, depression, autism, Parkinson's and Alzheimer's disease. Furthermore, alterations in zinc-levels have been associated with seizures and traumatic brain injury. Thus, altering zinclevels within the brain is emerging as a new target for the prevention and treatment of psychiatric and neurological diseases. However, given the restriction of zinc uptake into the brain by the blood-brain barrier, methods for controlled regulation and manipulation of zinc concentrations within the brain are rare. Here, we performed in vivo studies investigating the possibility of brain targeted zinc delivery using zinc-loaded nanoparticles which are able to cross the blood-brain barrier. After injecting these nanoparticles, we analyzed the regional and time-dependent distribution of zinc and nanoparticles within the brain. Moreover, we evaluated whether the presence of zinc-loaded nanoparticles alters the expression of zinc sensitive genes and proteins such as metallothioneins and zinc transporters and quantified possible toxic effects. Our results show that zinc loaded g7 nanoparticles offer a promising approach as a novel non - invasive method to selectively enrich zinc in the brain within a small amount of time.

Redistribution of labile plasma zinc during mild surgical stress in the rat.

Zinc is an essential trace element and cofactor for many cellular processes. Uptake of ionized divalent zinc (Zn(2+)) in peripheral tissues depends on its total content in the circulation and on mechanisms facilitating delivery to tissues in its labile form. Understanding mechanisms of Zn(2+) delivery has been hindered by the absence of techniques to detect labile Zn(2+) in the circulation. In this study, we report the use of the fluorescent zinc-binding dye (ZnAF-2) to detect changes in labile Zn(2+) in the circulating plasma of the rat under standardized conditions, including exogenous infusions to increase plasma Zn(2+) and an infusion of the chelator, citrate, to decrease labile Zn(2+) in the plasma without altering total Zn(2+) content. In a model of mild surgical stress (unilateral femoral arterial ligation), plasma levels of total and labile Zn(2+) decreased significantly 24 h after the operation. Ultrafiltration of plasma into high- and low-molecular weight macromolecule fractionations indicated that binding capacity of zinc in the high-molecular weight fraction is impaired for the entire 24-h interval after induction of mild surgical stress. Affinity of the filtrate fraction was rapidly and reversibly responsive to anesthesia alone, decreasing significantly at 4 h and recovering at 24 h; in animals subjected to moderate surgical stress, this responsiveness was lost. These findings are the first reported measurements of labile Zn(2+) in the circulation in any form of mild systemic stress. Zinc undergoes substantial redistribution in the plasma as a response to surgical stress, leading to increased availability in lower molecular weight fractions and in its labile form.

Coadministration of oral levofloxacin with agents that impair absorption: impact on antibiotic resistance.

Coadministration of oral divalent or trivalent cation-containing compounds with oral fluoroquinolones may impair fluoroquinolone absorption. Among 3,134 patients who received a course of oral levofloxacin, coadministration was significantly associated with subsequent identification of a levofloxacin-resistant isolate. Strategies to curb the emergence of fluoroquinolone resistance should include avoiding the coadministration of divalent or trivalent cation-containing compounds and fluoroquinolone.

[Magnesium ions prevent development of hyperkinesis produced by picrotoxin intrastriatal microinjections into the rat neostriatum].

The effects of daily bilateral microinjections of GABA-A receptor antagonist picrotoxin (2 mcg) into rostral neostriatum of the rats in chronic experiments were investigated. The picrotoxin was injected in volume of 1 mcl of saline or in 1 mcl of 1.0 M or 1.5 M MgCl2 solution; 1 mcl of saline or 1.0 M MgCl2 solution were injected in control groups of animals. The impairment of the avoidance conditioning in shuttle box, changes in free locomotor activity and the stereotypic choreo-myoclonic limb jerks were registered in rats with picrotoxin with the saline microinjections. The motor deviation registered are similar with Huntington chorea hyperkinesis in humans. The addition of magnesium ions into injected solution prevented both the lyperkinesis and condition behaviour realization impairment. As the magnesium is a universal calcium channel blocker, the calcium homeostasis of striatal neurons impairment as one of principal elements of hyperkinesis pathogenesis is proposed.

Coadministration of oral levofloxacin with agents that impair its absorption: potential impact on emergence of resistance.

Coadministration of fluoroquinolones (FQs) with divalent or trivalent cation-containing compounds (DTCCs) inhibits FQ absorption. In a case-control study of 46 inpatients receiving oral levofloxacin and DTCCs, patients with a levofloxacin-resistant isolate had been previously exposed to nearly twice as many days of levofloxacin/DTCC coadministration (P = 0.04). There remained a borderline significant independent association between the number of days of coadministration and levofloxacin-resistant culture [adjusted odds ratio (95% confidence interval) = 1.26 (0.98, 1.63); P = 0.07], even after controlling for the length of the levofloxacin course and the duration of hospitalisation prior to initiation of levofloxacin. Efforts should be directed at modifying hospital policies for dosing of levofloxacin and DTCCs to prevent coadministration.

Treatment failures secondary to drug interactions with divalent cations and fluoroquinolone.

We observed four cases of therapeutic failures while patients were simultaneously taking medications that contained divalent cations and oral fluoroquinolones. Patients improved after conversion to the intravenous formulation of the same antibiotics, proper spacing of the divalent cation, or conversion to a different antibiotic class. Patients prescribed oral fluoroquinolones should receive instructions on proper separations of these antibiotics with divalent cations. Health care professionals should be cognizant of these interactions and educated on their potential deleterious effect.

The Ca-activated Cl channel and its control in rat olfactory receptor neurons.

Odorants activate sensory transduction in olfactory receptor neurons (ORNs) via a cAMP-signaling cascade, which results in the opening of nonselective, cyclic nucleotide-gated (CNG) channels. The consequent Ca2+ influx through CNG channels activates Cl channels, which serve to amplify the transduction signal. We investigate here some general properties of this Ca-activated Cl channel in rat, as well as its functional interplay with the CNG channel, by using inside-out membrane patches excised from ORN dendritic knobs/cilia. At physiological concentrations of external divalent cations, the maximally activated Cl current was approximately 30 times as large as the CNG current. The Cl channels on an excised patch could be activated by Ca2+ flux through the CNG channels opened by cAMP. The magnitude of the Cl current depended on the strength of Ca buffering in the bath solution, suggesting that the CNG and Cl channels were probably not organized as constituents of a local transducisome complex. Likewise, Cl channels and the Na/Ca exchanger, which extrudes Ca2+, appear to be spatially segregated. Based on the theory of buffered Ca2+ diffusion, we determined the Ca2+ diffusion coefficient and calculated that the CNG and Cl channel densities on the membrane were approximately 8 and 62 micro m-2, respectively. These densities, together with the Ca2+ diffusion coefficient, demonstrate that a given Cl channel is activated by Ca2+ originating from multiple CNG channels, thus allowing low-noise amplification of the olfactory receptor current.

Ionized magnesium supplementation in critically ill patients: comparing ionized and total magnesium.

PURPOSE: The purpose of this study was to assess the effect of magnesium supplementation on total magnesium, ionized magnesium, ionized calcium, potassium, and pH in critically ill cancer patients and to compare the validity of the measurements. MATERIALS AND METHODS: Thirty-three consecutive critically ill patients receiving magnesium supplementation were placed in this prospective observational study at the Comprehensive Cancer Center, University Hospital. One gram (4.1 mmol) magnesium in 50 mL D5W was administered to critically ill patients, and the following were measured: total magnesium, ionized magnesium, ionized calcium, potassium, albumin, pH, BUN, creatinine, creatinine. RESULTS: Total magnesium and ionized magnesium increased by a mean of .11 +/- .02 and .05 +/- .01 mmol/L, respectively, after supplementation with 4.1 mmol of magnesium sulfate (P = .0001). Total magnesium, ionized magnesium, albumin, ionized calcium, potassium, and pH did not change significantly by the administration of 1 g of magnesium sulfate. CONCLUSION: The mean ionized magnesium (IMg+2) relationship to total magnesium (TMg) cannot be predicted before the supplementation with the available technology. After supplementation of 4.1 mmol/L the ionized magnesium level increased by .05 +/- .01 mmol/L. Magnesium supplementation had no significant effect on ionized calcium, potassium, and pH. TMg and IMg+2 should be followed independently.

Relaxant effect of magnesium and zinc on histamine-induced bronchoconstriction in dogs.

OBJECTIVE: Magnesium sulfate (MgSO4) has been reported to produce bronchodilation in asthmatic patients. In vitro studies have suggested that divalent cations inhibit L-type voltage-sensitive calcium ion (Ca2+) channels in cardiac and smooth muscles. In this study, we evaluated the in vitro and in vivo effects of magnesium ion (Mg2+) and zinc ion (Zn2+) on the airway contracted by histamine. SETTING: A university research laboratory. IN VITRO: Tracheal smooth muscle from guinea pigs. IN VIVO: Mongrel dogs. IN VITRO STUDY: The tension of isolated guinea pig tracheal strips was measured isometrically with a force displacement transducer. The specimen was contracted with histamine (10 microM). Then, MgSO4 (n = 6), zinc sulfate (ZnSO4, n = 6), or sodium sulfate (Na2SO4, n = 6) was cumulatively added to the organ bath. IN VIVO STUDY: The bronchial cross-sectional area of mongrel dogs was measured by a direct visualization method demonstrated previously. The dogs were randomly assigned to three groups: group Mg (n = 7), group Zn (n = 7), and group Na (n = 7). Bronchoconstriction was elicited with histamine (10 microg/kg plus 500 microg/kg/hr iv). Thirty minutes after the start of histamine infusion, 0 (saline), 1, 10, and 100 micromol/kg ZnSO4 or 1, 10, 100, and 1000 micromol/kg MgSO4 or Na2SO4 were administered intravenously in group Zn, Mg, or Na, respectively. The bronchial cross-sectional area was assessed before (basal) and 30 mins after the start of histamine infusion and 5 mins after each dose of ZnSO4, MgSO4, or Na2SO4. Arterial blood was also obtained to measure plasma levels of epinephrine and norepinephrine by gas chromatography-mass spectrometry. All data are expressed as mean +/- SEM. The doses of the divalent cations that reversed histamine-induced contraction by 50% were calculated by GraphPad Prism. MgSO4 and ZnSO4 (9.38+/-0.28 and 1.84+/-0.30 mM, respectively) relaxed histamine-contracted tracheal strip in a concentration-dependent manner, whereas Na2SO4 did not. Similarly, the in vivo study showed that MgSO4 and ZnSO4 dose-dependently reversed histamine-induced bronchoconstriction (potency, ZnSO4 > MgSO4), whereas Na2SO4 did not. In groups Mg and Zn, the plasma catecholamine levels also dose-dependently increased except when 1000 micromol/kg MgSO4 was administered. CONCLUSION: Because the divalent cations tested produced a spasmolytic effect on the contracted airway, infusion of divalent cations might be effective against asthmatic attack. However, high concentrations of these cations produce significant toxicity, so dosage will be an important concern in development of these agents.

The effect of extracellularly applied divalent cations on cytosolic Ca2+ in murine leydig cells: evidence for a Ca2+-sensing receptor.

1. The effect of extracellularly applied divalent cations upon cytosolic Ca2+ levels ([Ca2+]) was investigated in fura-2-loaded mouse Leydig (TM3) cells. 2. The extracellular application of Ca2+ (2.5-15 mM) or Ni2+ (0.5-5 mM) elicited concentration-dependent elevations in cytosolic [Ca2+] that were followed by decays to baseline levels. Extracellular Mg2+ (0.8-15 mM) failed to influence cytosolic [Ca2+]. 3. Conditioning applications of Ca2+ (2.5-10 mM), Mg2+ (2.5-15 mM) or Ni2+ (0.5-5 mM) all attenuated the cytosolic Ca2+ response to a subsequent test application of 5 mM [Ni2+]. 4. The amplitude of Ni2+-induced cytosolic Ca2+ signals remained constant in low-Ca2+ solutions. Such findings suggest a participation of Ca2+ release from intracellular stores. In parallel, depletion of Ca2+ stores by either ionomycin (5 microM, in low-Ca2+ solutions) or thapsigargin (4 microM) abolished or attenuated Ni2+-induced Ca2+ transients. 5. Ionomycin (5 microM) elevated cytosolic [Ca2+] in Ca2+-free solutions even after prior Ni2+ application, indicating the presence of Ni2+-insensitive stores. 6. Caffeine (250 and 500 microM) elevated cytosolic [Ca2+] and attenuated Ni2+-induced Ca2+ release. Furthermore, TM3 cells stained intensely with a specific anti-ryanodine receptor antiserum, Ab34. These findings suggest that Ca2+ release is regulated by ryanodine receptors. 7. Both membrane depolarization and hyperpolarization, brought about by changes in extracellular [K+] ([K+]e) in the presence of valinomycin (5 microM), altered the waveform of the Ni2+-induced cytosolic Ca2+ signal. Hyperpolarization, in addition, diminished the response magnitude. Such voltage-induced response modulation localizes the regulatory events to the Leydig cell plasma membrane. 8. We propose the existence of a cell surface divalent cation (Ca2+) receptor in Leydig cells, the activation of which triggers Ca2+ fluxes through ryanodine receptors.

Identification and characterisation of early reactions of asparagine-linked oligosaccharide assembly in Entamoeba histolytica.

Sequential incubation of a mixed membrane fraction isolated from Entamoeba histolytica trophozoites with the nonionic detergents Brij 35 and Igepal CA-630 rendered a soluble fraction with the ability to transfer N-acetylglucosamine (GlcNAc) from UDP-GlcNAc to dolichol phosphate to form a lipid saccharide that was identified as a mixture of dolichol-P-P-GlcNAc and dolichol-P-P-(GlcNAc)2 as follows. (a) The reaction occurred only in the presence of exogenously added dolichol phosphate and was strongly inhibited by tunicamycin and amphomycin; (b) Over 90% of the aminosugar moiety of the lipid saccharide was released by mild acid hydrolysis and was identified as a mixture of GlcNAc and diacetylchitobiose [(GlcNAc)2]; (c) Time course experiments revealed that dolichol-P-P-(GlcNAc)2 accumulated at the expense of a parallel decrease in dolichol-P-P-GlcNAc revealing the tandem operation of UDPGlcNAc:dolichol-P GlcNAc-1-P transferase and UDPGlcNAc:dolichol-P GlcNAc transferase. Mg2+ and to a lower extent Mn2+ were required for catalytic activity and were optimal at 2.5 mM and 1.25 mM, respectively. Common phospholipids with different head groups failed to increase catalytic activity and phosphatidylglycerol was inhibitory. At low concentration, nucleotides such as ATP, GMP and GTP brought about stimulations of 24-54% but higher concentrations were inhibitory. Others were inhibitory at all concentrations the strongest being those containing a uridine base.

Drug supplementation in pregnancy.

Since antiquity, there have been references in medicine to the role of nutrition in pregnancy outcome. Reviewing articles on nutrition and drug supplementation in pregnancy, one is struck by the variety of remedies that have been tried and the variety of effects that have been attributed to them. The number of herbal remedies that have been touted is astounding, and the entire science of Geophagia evolved in the hope identifying of those population-specific customs that may have had a positive effect on birth outcome as an adaptive mechanism. Most recently, there has been renewed interest in the role of nutritional and drug supplementation in pregnancy, specifically in the areas of pregnancy-induced hypertension and teratogenesis. In this article, I briefly review the role of drug supplementation in pregnancy, ranging from established needs such as iron to prevent iron-deficiency anemia to the controversies of low-dose aspirin supplementation for the prevention of preeclampsia and preconceptional folic acid supplementation for the prevention of neural tube defects.

Competitive action of divalent cations and D600 in frog slow muscle fibers.

Single, slow muscle fibers from Rana temporaria were equilibrated in normal Ringer's. 95 mmol/liter K(+)-solution containing various concentrations of Ca2+, Ni2+, Mn2+ or Mg2+ was applied, and the ensuing contractures were recorded isometrically. While peak tension (Fmax) was little affected, maintained tension (measured 1 min after onset of contracture) strongly depended on the concentration and species of divalent cations. Tension was maintained at its peak value in the presence of all species of divalent cations provided their concentrations were adequately increased. Dose-response curves were hyperbolic; Lineweaver-Burk plots revealed straight lines with different slopes intersecting near 1/Fmax, and indicating the following order of efficiency: Ni2+ greater than Ca2+ greater than Mn2+ much greater than Mg2+. Hill plots for these cations resulted in straight lines with slopes near 1. Qualitatively similar relationships were obtained with contracture solutions containing D600 (3-12 mumol/liter). However, under these conditions higher concentrations of Ca2+ or Ni2+ were required in order to fully maintain tension. After a step concentration change in the medium during contracture, the effects of Ca2+ or D600 were detectable only after a delay of 9 and 18 sec, respectively. It is concluded that divalent cations and D600 compete for the same binding site according to a 1:1 reaction. This site is presumably located inside the transverse tubular system and controls inactivation of the contractile force.

Effect of liposomes containing various divalent cations on the release of acetylcholine from synaptosomes.

The effect of increasing the cytoplasmic levels of various divalent cations on the release of [3H]acetylcholine ([3H]ACh) from synaptosomes was investigated. Synaptosomes prepared from rat brain and prelabeled with [3H]choline were incubated with liposomes containing Mg2+, Ca2+, Mn2+, Co2+, Sr2+, or Ba2+. This treatment allows the transfer of the aqueous contents of the liposomes to the cytoplasm of the synaptosomes. The efflux of radioactivity subsequent to this treatment was measured, and the relative proportions of [3H]ACh and [3H]choline were determined. The release of radioactivity from synaptosomes incubated with liposomes containing Mg2+, Mn2+, or Co2+ was not altered when compared with synaptosomes incubated either without liposomes or with liposomes containing isotonic K+/Na+. Synaptosomes incubated with liposomes containing Ca2+, Sr2+, or Ba2+, however, released significantly more radioactivity than did controls. Moreover, the released radioactivity consisted almost entirely of [3H]ACh. Liposomes containing either Ca2+ or Sr2+ were equally effective in promoting the release of [3H]ACh from synaptosomes, whereas liposomes containing Ba2+ were 2.5 times more effective in promoting the release of [3H]ACh than were liposomes containing either Ca2+ or Sr2+. Since liposomes introduce their aqueous contents into cytoplasm via a mechanism not involving plasma membrane channels, the increased release of [3H]ACh caused by liposomes containing Ca2+, Sr2+, or Ba2+ is attributable to an increase in the intrasynaptosomal concentration of these ions, and not to their passage through calcium channels.