Spray Dried Smectite Clay Particles as a Novel Treatment against Obesity.

PURPOSE: To explore the feasibility of spray dried smectite clay particles fabricated from montmorillonite or laponite materials for adsorbing dietary lipids and reducing rodent weight gain in vivo. METHODS: Spray dried montmorillonite (SD-MMT) and spray dried laponite (SD-LAP) particles were prepared via spray drying. Particle morphology, surface area and redispersion/aggregation properties in aqueous media were characterized. The ability of SD-MMT and SD-LAP particles to inhibit lipid digestion kinetics and adsorb lipid species from solution was assessed during in vitro lipolysis using proton nuclear magnetic resonance analysis. SD-MMT and SD-LAP particles were dosed to rodents fed a high-fat diet and their effect on body weight gain was evaluated. RESULTS: Both SD-MMT and SD-LAP particles adsorbed significant quantities of medium chain triglycerides and lipolytic products from solution during in vitro lipolysis. At a concentration of 50% w/w relative to lipid content, SD-MMT and SD-LAP particles adsorbed 42% and 94% of all lipid species, respectively. SD-MMT and SD-LAP particles also reduced the extent of rodent weight gain relative to the negative control treatment group and performed similarly to orlistat via an alternate mechanism of action. CONCLUSIONS: Spray dried smectite clay particles (SD-MMT and SD-LAP) with significant adsorptive capacities for dietary lipids and digestion products were successfully fabricated. These particles may be developed as novel anti-obesity treatments with fewer adverse effects than currently marketed treatment options.

Aripiprazole-montmorillonite: a new organic-inorganic nanohybrid material for biomedical applications.

Poor aqueous solubility and the unpleasant taste of aripiprazole (APZ) have been recurring problems, owing to its low bioavailability and low patient tolerance, respectively. Herein, we prepared a nanohybrid system that was based on a bentonite clay material, montmorillonite (MMT), which could both mask the taste and enhance the solubility of APZ (i.e., APZ-MMT). To further improve the efficacy of this taste masking and drug solubility, APZ-MMT was also coated with a cationic polymer, polyvinylacetal diethylamino acetate (AEA). In vitro dissolution tests at neutral pH showed that the amount of drug that was released from the AEA-coated APZ-MMT was greatly suppressed (<1%) for the first 3 min, thus suggesting that AEA-coated APZ-MMT has strong potential for the taste masking of APZ. Notably, in simulated gastric juice at pH 1.2, the total percentage of APZ that was released within the first 2 h increased up to 95% for AEA-coated APZ-MMT. Furthermore, this in vitro release profile was also similar to that of Abilify®, a commercially available medication. In vivo experiments by using Sprague-Dawley rats were also performed to compare the pharmacokinetics of AEA-coated APZ-MMT and Abilify®. AEA-coated APZ-MMT exhibited about 20% higher systemic exposure of APZ and its metabolite, dehydro-APZ, compared with Abilify®. Therefore, a new MMT-based nanovehicle, which is coated with a cationic polymer, can act as a promising delivery system for both taste masking and for enhancing the bioavailability of APZ.

Development of an oral bentonite-based modified-release freeze-dried powder of vactosertib: Pharmacokinetics and anti-colitis activity in rodent models of ulcerative colitis.

Vactosertib is a novel inhibitor of transforming growth factor-ß signaling. Clinical applications of vactosertib have been challenging since conventional oral formulations such as immediate-release tablets demonstrate a rapid rise and fast decline in plasma concentrations. In this study, a novel bentonite-based, modified-release, freeze-dried powder of vactosertib was developed and evaluated to determine its potential in the treatment of ulcerative colitis. The formulation released vactosertib slowly and steadily in an in vitro drug release test. The extent of vactosertib released from the formulation was markedly low (18.0%) at pH 1.2 but considerably high (95.6%) at pH 7.4. Compared with vactosertib oral solution, the formulation demonstrated a 52.5% lower mean maximum concentration of vactosertib and three times longer median time to maximum concentration without a significant change in the extent of vactosertib absorption in a rodent colitis model. Furthermore, colitis mice administered with this formulation showed a significant reduction in the total histopathological score by 30% compared with those administered with the positive control, whereas the administration of vactosertib oral solution resulted in only a 10% reduction. Collectively, this novel formulation resolved the pharmacokinetic drawbacks of vactosertib and is expected to enhance its therapeutic effect by delivering vactosertib to the colitis lesions in the lower gastrointestinal tract.

Artificial model of photosynthetic oxygen evolving complex: catalytic O2 production from water by di-mu-oxo manganese dimers supported by clay compounds.

Adsorption of [(OH(2))(terpy)Mn(mu-O)(2)Mn(terpy)(OH(2))](3+) (terpy=2,2':6',2"-terpyridine) (1) onto montmorillonite K10 (MK10) yielded catalytic dioxygen (O(2)) evolution from water using a Ce(IV) oxidant. The Mn K-edge X-ray absorption near edge structure (XANES) of the 1/MK10 hybrid suggested that the oxidation state of the di-mu-oxo Mn(2) core could be Mn(III)-Mn(IV). However the pre-edge peak in the XANES spectrum of 1 adsorbed on MK10 is different from the neat 1 powder. The kinetic analysis of O(2) evolution showed that the catalysis requires cooperation of two equivalents of 1 adsorbed on MK10. The reaction of the [(bpy)(2)Mn(mu-O)(2)Mn(bpy)(2)](3+) (bpy=2,2'-bipyridine) (2)/MK10 hybrid with a Ce(IV) oxidant evolved O(2). However, the turnover number value was less than unity for 2/MK10, showing that 2 adsorbed on MK10 does not work as a catalyst. The terminal water ligands could be an important for the catalysis by adsorbed 1. The mechanism of O(2) production by photosynthetic oxygen evolving complex is discussed based on catalytic O(2) evolution by 1 adsorbed on MK10.

Interaction of Pseudomonas putida with kaolinite and montmorillonite: a combination study by equilibrium adsorption, ITC, SEM and FTIR.

Equilibrium adsorption along with isothermal titration calorimetry (ITC), Fourier transform infrared spectra (FTIR) and scanning electron microscopy (SEM) techniques were employed to investigate the adsorption of Pseudomonas putida on kaolinite and montmorillonite. A higher affinity as well as larger amounts of adsorption of P. putida was found on kaolinite. The majority of sorbed bacterial cells (88.7%) could be released by water from montmorillonite, while only a small proportion (9.3%) of bacteria desorbed from kaolinite surface. More bacterial cells were observed to form aggregates with kaolinite, while fewer cells were within the larger bacteria-montmorillonite particles. The sorption of bacteria on kaolinite was enthalpically more favorable than that on montmorillonite. Based on our findings, it is proposed that the non-electrostatic forces other than electrostatic force play a more important role in bacterial adsorption by kaolinite and montmorillonite. Adsorption of bacteria on clay minerals resulted in obvious shifts of infrared absorption bands of water molecules, showing the importance of hydrogen bonding in bacteria-clay mineral adsorption. The enthalpies of -4.1+/-2.1 x 10(-8) and -2.5+/-1.4 x 10(-8)mJ cell(-1) for the adsorption of bacteria on kaolinite and montmorillonite, respectively, at 25 degrees C and pH 7.0 were firstly reported in this paper. The enthalpy of bacteria-mineral adsorption was higher than that reported previously for bacteria-biomolecule interaction but lower than that of bacterial coaggregation. The bacteria-mineral adsorption enthalpies increased at higher temperature, suggesting that the enthalpy-entropy compensation mechanism could be involved in the adsorption of P. putida on clay minerals. Data obtained in this study would provide valuable information for a better understanding of the mechanisms of mineral-microorganism interactions in soil and associated environments.

PEG-PE/clay composite carriers for doxorubicin: Effect of composite structure on release, cell interaction and cytotoxicity.

A novel drug delivery system for doxorubicin (DOX), based on organic-inorganic composites was developed. DOX was incorporated in micelles (M-DOX) of polyethylene glycol-phosphatidylethanolamine (PEG-PE) which in turn were adsorbed by the clay, montmorillonite (MMT). The nano-structures of the PEG-PE/MMT composites of LOW and HIGH polymer loadings were characterized by XRD, TGA, FTIR, size (DLS) and zeta measurements. These measurements suggest that for the LOW composite a single layer of polymer intercalates in the clay platelets and the polymer only partially covers the external surface, while for the HIGH composite two layers of polymer intercalate and a bilayer may form on the external surface. These nanostructures have a direct effect on formulation stability and on the rate of DOX release. The release rate was reversely correlated with the degree of DOX interaction with the clay and followed the sequence: M-DOX>HIGH formulation>LOW formulation>DOX/MMT. Despite the slower release from the HIGH formulation, its cytotoxicity effect on sensitive cells was as high as the "free" DOX. Surprisingly, the LOW formulation, with the slowest release, demonstrated the highest cytotoxicity in the case of Adriamycin (ADR) resistant cells. Confocal microscopy images and association tests provided an insight into the contribution of formulation-cell interactions vs. the contribution of DOX release rate. Internalization of the formulations was suggested as a mechanism that increases DOX efficiency, particularly in the ADR resistant cell line. The employment of organic-inorganic hybrid materials as drug delivery systems, has not reached its full potential, however, its functionality as an efficient tunable release system was demonstrated. STATEMENT OF SIGNIFICANCE: DOX PEG-PE/clay formulations were design as an efficient drug delivery system. The main aim was to develop PEG-PE/clay formulations of different structures based on various PEG-PE/clay ratios in order to achieve tunable release rates, to control the external surface characteristics and formulation stability. The formulations showed significantly higher toxicity in comparison to "free" DOX, explained by formulation internalization. For each cell line tested, sensitive and ADR resistant, a different formulation structure was found most efficient. The potential of PEG-PE/clay-DOX formulations to improve DOX administration efficacy was demonstrated and should be further explored and implemented for other cancer drugs and cells.

Fate of prions in soil: trapped conformation of full-length ovine prion protein induced by adsorption on clays.

Studying the mechanism of retention of ovine prion protein in soils will tackle the environmental aspect of potential dissemination of scrapie infectious agent. We consider the surface-induced conformational changes that the recombinant ovine prion protein (ovPrP) may undergo under different pH conditions when interacting with soil minerals of highly adsorptive capacities such as montmorillonite. The conformational states of the full-length ovine prion protein adsorbed on the electronegative clay surface are compared to its solvated state in deuterated buffer in the pD range 3.5-9, using FTIR spectroscopy. The in vitro pH-induced conversion of the alpha-helical monomer of ovPrP into oligomers of beta-like structure prone to self-aggregation does not occur when the protein is adsorbed on the clay surface. The conformation of the trapped ovPrP molecules on montmorillonite is pH-independent and looks like that of the ovPrP solvated state at pD higher than 7, suggesting the major role of Arg and Lys residues in the electrostatic origin of adsorption. The uneven distribution of positively and negatively charged residues of the ovPrP protein would promote a favored orientation of the protein towards the clay, so that not only the basic residues embedded in the N-terminal flexible part but also external basic residues in the globular part of the protein might participate to the attractive interaction. From these results, it appears unlikely that the interaction of normal prions (PrP(C)) with soil clay surfaces could induce a change of conformation leading to the pathogenic form of prions (PrP(Sc)).

Antibacterial effects of the Cu(II)-exchanged montmorillonite on Escherichia coli K88 and Salmonella choleraesuis.

The aim of this research was to determine the antibacterial properties and mechanisms of Cu(II)-exchanged montmorillonite (MMT-Cu) in vitro. Escherichia coli ATCC K88 and Salmonella choleraesuis ATCC 50020 were chosen as indicators of intestinal tract pathogenic bacteria in weanling pigs. The antibacterial activity of MMT-Cu and MMT were evaluated by determining the minimum inhibitory concentrations (MICs) using two-fold serial dilutions in MH broth, and the amount of Cu2+ released into the broth was measured by an atomic absorption technique. The rate of oxygen consumption was measured using a SP-II-type oxygen electrode analyzer; the structural integrity of cell walls of bacteria was observed by transmission electron microscope (TEM); enzymatic activity of bacteria was examined with a semi-automatic biochemical analyzer. The results showed that MMT-Cu inhibited the growth of E. coli K88 and S. choleraesuis, and the MICs were 1024 and 2048 microg/ml, respectively. The amount of Cu2+ released into the broth was in the range 6.51-45.65 microg/ml. Nevertheless, both tested bacteria still grew in broth containing 32,768 microg/ml of MMT. Treatment with MMT-Cu could lead to significant release of intracellular enzymes from the tested bacteria. Data from oxygen consumption of bacteria showed that MMT-Cu could inhibit the TCA pathway of the bacterial respiration metabolism. These results show that MMT-Cu has an antibacterial activity.

The toxicity of insoluble cerium-144 inhaled by beagle dogs: non-neoplastic effects.

The biological effects of inhaled beta-particle-emitting radionuclides are not well known. The non-neoplastic diseases induced by an inhaled, relatively insoluble form of cerium-144 ((144)Ce) were studied in beagle dogs exposed to graded activity levels of (144)Ce in fused aluminosilicate particles by a single, brief inhalation exposure and observed for their life span. The initial lung burdens (ILBs) achieved ranged from 0.000093-7.6 MBq (144)Ce/kg body weight. The (144)Ce was retained in the lung with an effective half-life of about 190 days. Significant (144)Ce was translocated to the tracheobronchial lymph nodes, and the concentration exceeded that of the lung at about 400 days after inhalation exposure. Significant radiation doses were delivered to the lung and tracheobronchial lymph nodes and to the heart adjacent to the tracheobronchial lymph nodes. Radiation pneumonitis was the predominant non-neoplastic disease. The dose response for radiation pneumonitis indicated that an ILB of 1.4 MBq/kg would cause death from radiation pneumonitis in 50% of the exposed dogs. This ILB resulted in a pulmonary dose to death of about 350 Gy. The tracheobronchial lymph nodes developed lesions in dogs with ILBs lower than those causing radiation pneumonitis. The overall results of this study, however, showed that (144)Ce, inhaled in an insoluble form, did not cause any unique or inexplicable biological effects in dogs or cause effects at unusually low doses that might call current radiation protection guidelines into question.

Effect of the different factors on the iontophoretic delivery of calcium ions from bentonite.

The calcium content of mud patches used for therapy is very small. Several mineral clays originating from Hungary served as a base material for experiments in order to find a suitable drug for transdermal introduction of calcium ions into the body. The Ca++ transport through the pig skin has been investigated in vitro in diffusion cells applying iontophoresis. Studies of electrical and physicochemical factors acting on the permeation kinetics of in vitro experiments were performed. The utilization of direct current has intensified the Ca++ transport through the pig skin (129.78+/-26. 15 microgram Ca/cm2). On using pulsate currents the amount of the Ca++ penetrating through the skin was 5-10 times higher (283.18+/-16.89 microgram Ca/cm2, 388.71+/-19.90 microgram Ca/cm2) than that of the passive transport (36.22+/-14.20 microgram Ca/cm2). The amount of Ca++ cumulated in the receptor compartment was directly proportional to the amount of bentonite (a natural mineral clay with a large cation exchange capacity) in the donor compartment and to the concentration of Ca++ in the lattice of the applied mineral clay. Therefore, the experiments were carried out on a bentonite previously enriched in Ca++ in its lattice (50 mg Ca/g bentonite). The results of the in vitro studies could open a new field of application in the therapy of osteoporosis or in the use of mineral substances.

Interaction between tylosin and bentonite clay from a pharmacokinetic perspective.

The interaction between bentonite and tylosin was investigated in broiler chickens, based on pharmacokinetic characteristics obtained in vivo. Simultaneous oral administration of bentonite and tylosin significantly lowered plasma levels of tylosin and reduced the area under the plasma concentration-time curve (AUC(0-inf)), maximal plasma concentration (C(max)), time to maximal plasma concentration (T(max)) and relative oral bioavailability. The results prove unambiguously the binding of tylosin by bentonite. Simultaneous administration of tylosin (in the drinking water or feed) and bentonite (mixed in the feed as a mycotoxin binder) should therefore be avoided.

Montmorillonite intercalated with glutathione for antioxidant delivery: synthesis, characterization, and bioavailability evaluation.

A most powerful antioxidant, glutathione (GSH), plays an important role in detoxification, immune response, and protection against reactive oxygen species. However, orally ingested GSH can be easily degradable to free amino acids by chemical and enzymatic hydrolysis, resulting in low bioavailability. The aim of this study was, therefore, to enhance GSH bioavailability by developing GSH-montmorillonite (MMT) hybrid system. It was also coated with polyvinylacetal diethylaminoacetate (AEA) for better stability. Both GSH-MMT and AEA-GSH-MMT hybrids were characterized by powder X-ray diffraction (PXRD), Fourier transformed infrared (FT-IR), and thermogravimetric analysis (TGA), indicating that GSH was successfully intercalated into the interlayer spaces of MMT. In vivo antioxidant activity assay revealed that AEA-GSH-MMT hybrid significantly increased antioxidant activity in the plasma after oral administration in mice. Pharmacokinetic study also indicated that AEA-GSH-MMT hybrid considerably increased the plasma concentration of GSH at 1h post-oral administration. Moreover, both the hybrid systems remarkably enhanced GSH delivery to the main target tissue, liver. All the results suggest that GSH-MMT hybrid systems have great potential to enhance bioavailability of oral GSH, providing new insight into their pharmaceutical application.

Production, characterization and (co-)immobilization of dextranase from Penicillium aculeatum.

Fermentation kinetics of Penicillium aculeatum ATCC 10409 demonstrated that fungal growth and dextranase release are decoupled. Inoculation by conidia or mycelia resulted in identical kinetics. Two new isoenzymes of the dextranase were characterized regarding their kinetic constants, pI, MW, activation energy and stabilities. The larger enzyme was 3-fold more active (turnover number: 2,230 +/- 97 s(-1)). Pre-treatment of bentonite with H(2)O(2) did not affect adsorption characteristics of dextranase. Enzyme to bentonite ratios above 0.5:1 (w/w) resulted in a high conservation of activity upon adsorption. Furthermore, dextranase could be used in co-immobilizates for the direct conversion of sucrose into isomalto-oligosaccharides (e.g. isomaltose). Yields of co-immobilizates were 2-20 times that of basic immobilizates, which consist of dextransucrase without dextranase.

Desempenho de perus de corte alimentados com níveis crescentes de afloxinas, com ou sem adiçäo de adsorvente / Turkey performance fed with increasing aflatoxins levels, with or without adsorvent inclusion

Este trabalho foi realizado durante o veräo de 1995/1996 com o objetivo de determinar o nível de aflatoxinas (AFL) capaz de causar prejuízos no desempenho de perus de corte criados sob condiçöes ambientais de baixo desafio, bem como avaliar a eficácia da adiçäo de bentonita sódica natural (BSN) como um adsorvente. Foram utilizados 1008 perus de corte, alojados em 84 boxes seguindo um delineamento experimental de blocos casualizados com 14 tratamentos distribuídos em um arranjo fatorial 7 x 2, sendo 7 níveis de adiçäo de AFL (0, 10, 50, 100, 500, 1000 e 2000ppb) e 2 níveis de adiçäo de BSN (0 e 0,5 porcento). Aos 21 dias de idade, as aves alimentadas com as dietas sem adiçäo de BSN mostraram uma reduçäo significativa (P<0,05) no ganho de peso (GP) e no consumo de raçäo (CR) com níveis de AFL iguais ou superiores a 500ppb, enquanto que aves alimentadas com as dietas com 0,5 porcento de BSN mostraram reduçäo significativa (P<0,05) no GP e CR apenas à partir de 1000ppb de AFL. Estes mesmos resultados foram observados aos 40 dias de idade, entretanto aos 70 dias ocorreu uma reduçäo significativa (P<0,05) no GP e CR das aves à partir de 500ppb de AFL, com ou sem adiçäo de BSN, demonstrando um efeito cumulativo desta micotoxina. Em geral, em todos os níveis de adiçäo de AFL que ocorreu reduçäo significativa no GP e CR das aves, esta foi cerca de 15 a 20 porcento menor nos grupos alimentados com 0,5 porcento de BSN. A conversäo alimentar (CA) das aves foi menos afetada pelos níveis de AFL dietéticos, embora tenha existido uma tendência de piora na CA com altos níveis de AFL, em todas as idades. Aflatoxinas tiveram um grande efeito sobre a taxa de mortalidade (porcentagem MOR), sendo que já aos 21 dias a porcentagem MOR verificada nos lotes que receberam 1000 e 2000ppb de AFL, sem BSN, foi de 15,3 e 72,2 porcento, respectivamente. Antes de completar 40 dias de experimento, 100 porcento das aves alimentadas com dietas contendo 2000ppb de AFL, sem e com BSN, morreram. A adiçäo de 0,5 porcento de BSN proporcionaou uma reduçäo média de 35 porcento na porcentagem MOR em comparaçäo aves alimentadas com dietas sem BSN. Com base nos resultados pode ser concluído que AFL säo extremamente deletérias aos perus e que a adiçäo de 0,5 porcento de BSN na raçäo protege particalmente os perus dos efeitos negativos desta micotoxina.

Terapias alternativas; la Montmorillonita-bentonita como antiacido

La farmacopea de las culturas nativas bolivianas contienen productos con actividad farmacologica, como es el caso de la Montmorillonita-bentonita (Phasa), arcilla que a traves de analisis quimicos se ha demostrado que contiene silicatos, hidroxido y carbonatos, principalmente de aluminio y calcio analoga a los tradicionales antiacidos del arsenal terapeutico facultativo.