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Preparation, characterization and protein loading of hexanoyl-modified chitosan nanoparticles.

Hexanoyl chitosan was synthesized through a coupling reaction between chitosan and hexanoic anhydride. Proton nuclear magnetic resonance (1HNMR) and fourier-transform infrared (FTIR) spectroscopy studies showed the formation of hexanoyl chitosan. The nanoparticles of hexanoyl chitosan were prepared through ionotropic gelation with tripolyphosphate (TPP) followed by sonication. The hexanoyl chitosan-TPP nanoparticles exhibited uniform spherical shape with smooth surface as observed by atomic force microscopy and transmission electron microscopy. The particle size of nanoparticles was between 54.1 to 724 nm with a mean diameter of 324 nm. At 0.2, 0.4, and 0.6 mg/mL bovine serum albumin initial concentration, the encapsulation efficiency and loading capacity of hexanoyl-chitosan-TPP nanoparticles were 58.2, 44.5, and 28.1%, and 14.1, 23.4, and 30.3%, respectively.

Caproates↗

Composition of the essential oil of rootstock from Cimicifuga simplex.

The composition of the essential oils from rootstock of Cimicifuga simplex has been investigated by capillary GC and GC/MS. The main components in essential oil were m-acetanisole (27.57%), (E)-cinnamaldehyde (6.84%), paeonol (5.58%), caproic acid (5.07%) and atractylone (3.10%). The oil was characterized by a high content of aromatic components (52.59%).

Acetophenones↗

Isolation and synthesis of acmellonate, a new unsaturated long chain 2-ketol ester from Spilanthes acmella.

Paracress (Spilanthes acmella) is commonly used as a spice, e.g., in Madagascar. To get more information about the taste active compounds, Jambu oleoresin produced from Spilanthes acmella and extracts prepared directly from the plant were fractionated and the fractions were analysed and evaluated by tasting. In addition to the long known tingling compounds (2E,6Z,8E)-deca-2,6,8-trienoic acid N-isobutyl amide (spilanthol) and (2E,6Z,8E)-deca-2,6,8-trienoic acid N-(2-methylbutyl) amide, as a minor constituent also the new 2-ketol ester (7Z,9E)-2-oxo-undeca-7,9-dienyl 3-methylbut-2-enoate (acmellonate) was isolated. The structure of the latter compound was verified by total synthesis of the ketol ester via the reaction of tris(trimethylsilyloxy)ethene and (6Z,8E)-deca-6,8-dienoic acid chloride and subsequent classical esterification. Acmellonate shows a weak tingling and numbing effect on the tongue and contributes only to a small extend to the overall flavour of Jambu.

Asteraceae↗

Biliary excretion of [14C]penicillic acid by rats.

Adult male rats were given [14C]penicillic acid by oral intubation, and the biliary excretion of the compound was monitored. The significance of bile as an excretory route was confirmed and compared by using open and recycle cannulation of the bile duct. The biological half-retention life of [14C]penicillic acid in the bile was 3.63 h.

Animals↗

Effect of penicillic acid on biliary excretion of indocyanine green in the mouse and rat.

Penicillic acid (PA), a mycotoxin, is hepatotoxic. A study was undertaken to investigate its effects on hepatobiliary excretory function, using the anionic compounds indocyanine green (ICG), in mice and rats. Pretreatment with a single dose of PA (90 mg/kg, ip, an LD50 dose in both species) resulted in depression of ICG excretion in both species. This depression was dose- and time-dependent. Decreases of 42 and 57% in biliary excretion of ICG were observed in rats and mice 48 and 72 h after PA pretreatment, respectively. Although bile flow was depressed significantly when expressed in terms of body weight, it was not altered in mice when expressed in terms of liver weight. Bile flow was not affected in rats. While the serum ICG concentration was increased after PA treatment in both species, the liver ICG concentration was not affected. The liver-to-serum, bile-to-serum, and bile-to-liver ICG concentration ratios decreased in PA-treated animals. These data suggest that the PA-induced hepatobiliary excretory dysfunction may result from depression of both uptake of ICG into the liver and bile canlicular transport of ICG.

Animals↗

Evaluation of gas-liquid chromatography (GLC) for rapid detection of Clostridium difficile in fecal specimens.

Clostridium difficile intestinal infection is a major nosocomial hazard in patients receiving antimicrobial therapy. Rationale for rapid diagnosis include lifesaving antimicrobial therapy in patients with severe colitis and early isolation measures for transmission control. We have therefore analysed the sensitivity, specificity and predictive value of GLC identification of isocaproic acid in diarrheic stools from adult hospitalized patients in comparison with selective fecal culture on Cycloserine Cefoxitin Fructose Agar. During the study period, the prevalence of positive culture for C. difficile was 38/595 fecal specimens (6.4%). Compared with culture, GLC had a sensitivity of 24/38 (63%) and a specificity of 524/557 (94%). The predictive value of a positive GLC was 24/57 (42%) and of a negative GLC was 524/538 (97%). Measurement of the isocaproic acid peak height did not allow determination of a cutt-off value improving the test accuracy. The sensitivity of detection of isocaproic acid in stools by GLC is too low to be used as screening test for C. difficile infection. However, in a low prevalence population, a positive GLC test increased the pre-test probability of infection sevenfold.

Adult↗

Stability and in vitro drug release of flurbiprofen-loaded poly-epsilon-caprolactone nanospheres.

The effects of temperature and two different initial pH (2.67 and 7.00) on poly-epsilon-caprolactone (P epsilon CL) nanospheres loaded with flurbiprofen (FB) (aqueous suspensions) were studied to investigate their influence on the stability and physicochemical characteristics of these drug delivery systems. The drug release behavior was also studied. Release of the associated FB occurred very fast on high dilution in a buffered medium. The stability of the polymeric system depends on the temperature and the initial pH value; it is more degradable with the particles stored at 40 degrees C with an initial pH value of 2.67.

Administration, Topical↗

Destruction of epidermal cells in vitro by autologous serum from normal animals.

Sera and plasma from normal rats and rabbits were shown to be extremely toxic in vitro to autologous epidermal cells. On the other hand, mouse sera and newborn rat sera were innocuous to autologous epidermal cells. Viability of cells was assessed by the method of eosin dye exclusion upon 2 hour incubation at 37 degrees C. Testicular cells were also killed by autologous sera, but polymorphonuclear leukocytes, lymphocytes, and lymph node cells were not affected. Autotoxicity of sera could be destroyed by the depletion of complement components with an antigen-antibody precipitate, heat, zymosan, and NH(3). Moreover, activity of sera could be absorbed out by epidermal cells, though not by lymph node cells or erythrocytes. Such absorption of toxicity was not individual-specific since homologous epidermal cells also absorbed toxicity, and in addition, were killed by fresh normal serum. Enzyme inhibitors such as soybean trypsin inhibitor and epsilon-amino-n-caproic acid did not affect the activity of fresh autologous serum. It is suggested that a natural barrier exists between the basal cells of the epidermis and the plasma which prevents the autodestructive process under normal conditions. Any injury to this barrier may than lead to necrosis and death of the epidermis as seen in various pathological conditions.

Animals↗

Water and nonelectrolyte permeability of lipid bilayer membranes.

Both the permeability coefficients (Pd's) through lipid bilayer membranes of varying composition (lecithin [L], lecithin:cholesterol [LC], and spingomyelin:cholesterol [SC]) and the n-hexadecane:water partition coefficients (Knc's) of H2O and seven nonelectrolytes (1,6 hexanediol, 1,4 butanediol, n-butyramide, isobutyramide, acetamide, formamide, and urea) were measured. For a given membrane compositiin, Pd/DKnc (where D is the diffusion constant in water) is the same for most of the molecules tested. There is no extraordinary dependence of Pd on molecular weight; thus, given Pd(acetamide), Pd(1,6 hexanediol) is correctly predicted from the Knc and D values for the two molecules. The major exceptions are H2O, whose value of Pd/DKnc is about 10-fold larger, and urea, whose value is about 5-fold smaller than the general average. In a "tight" membrane such as SC, Pd(n-butyramide)/Pd(isobutyramide)=2.5; thus this bilayer manifests the same sort of discrimination between branched and straight chain molecules as occurs in many plasma membranes. Although the absolute values of the Pd's change by more than a factor of 100 in going from the tightest membrane (SC) to the loosest (L), the relative values remain approximately constant. The general conclusion of this study is that H2O and nonelectrolytes cross lipid bilayer membranes by a solubility-diffusion mechanism, and that the bilayer interior is much more like an oil (a la Overton) than a rubber-like polymer (a la Lieb and Stein).

Acetamides↗

Intestinal transport of weak electrolytes: Determinants of influx at the luminal surface.

The determinants of weak electrolyte influx into everted segments of rat small intestine have been studied. Preliminary experiments showed that the observed influxes could be described as unidirectional, diffusional fluxes of the nonionized compound uncomplicated by a parallel ionic component. It is shown that the determinants of weak electrolyte influx in this situation may be described in terms of the resistance of the unstirred layer to movement from the bulk phase to the cell surface, the degree of ionization of the weak electrolyte at the cell surface, and the cellular permeability to the nonionized weak electrolyte. Quantitative considerations indicated that the unstirred layer was totally rate-limiting in the cases of some poorly ionized, or highly permeant compounds, but the unstirred layer was not totally rate limiting for most of the compounds studied. Calculation of cellular permeabilities for the nonionized forms of weak electrolytes required assumptions to be made concerning the pH value in the surface fluid layer. A uniform set of permeability data including both weak acids and weak bases was obtained only when it was assumed that the pH in the surface fluid layer was equal to that in the bulk phase, and it was concluded that these studies do not support the concept of a microclimate of distinctive pH at the epithelial surface as a determinant of weak electrolyte transport.

Animals↗

Osteogenic induction of human bone marrow-derived mesenchymal progenitor cells in novel synthetic polymer-hydrogel matrices.

The aim of this project was to investigate the in vitro osteogenic potential of human mesenchymal progenitor cells in novel matrix architectures built by means of a three-dimensional bioresorbable synthetic framework in combination with a hydrogel. Human mesenchymal progenitor cells (hMPCs) were isolated from a human bone marrow aspirate by gradient centrifugation. Before in vitro engineering of scaffold-hMPC constructs, the adipogenic and osteogenic differentiation potential was demonstrated by staining of neutral lipids and induction of bone-specific proteins, respectively. After expansion in monolayer cultures, the cells were enzymatically detached and then seeded in combination with a hydrogel into polycaprolactone (PCL) and polycaprolactone-hydroxyapatite (PCL-HA) frameworks. This scaffold design concept is characterized by novel matrix architecture, good mechanical properties, and slow degradation kinetics of the framework and a biomimetic milieu for cell delivery and proliferation. To induce osteogenic differentiation, the specimens were cultured in an osteogenic cell culture medium and were maintained in vitro for 6 weeks. Cellular distribution and viability within three-dimensional hMPC bone grafts were documented by scanning electron microscopy, cell metabolism assays, and confocal laser microscopy. Secretion of the osteogenic marker molecules type I procollagen and osteocalcin was analyzed by semiquantitative immunocytochemistry assays. Alkaline phosphatase activity was visualized by p-nitrophenyl phosphate substrate reaction. During osteogenic stimulation, hMPCs proliferated toward and onto the PCL and PCL-HA scaffold surfaces and metabolic activity increased, reaching a plateau by day 15. The temporal pattern of bone-related marker molecules produced by in vitro tissue-engineered scaffold-cell constructs revealed that hMPCs differentiated better within the biomimetic matrix architecture along the osteogenic lineage.

Alkaline Phosphatase↗

Tissue biocompatibility of a new caprolactone-coated self-reinforced self-expandable poly-L-lactic acid bioabsorbable urethral stent.

BACKGROUND AND PURPOSE: The bioabsorption time as well as other properties of bioabsorbable polymers can be affected by the choice of the basic molecule, by the degree of its polymerization, and by the coating material used in the device. The aim of our present study was to evaluate the biocompatibility of a new caprolactone copolymer-coated, self-reinforced poly-L-lactic acid (SR-PLLA) urethral stent by means of a rabbit muscle implantation test. This new material has previously been tested for cytotoxicity using the thymidine incorporation method (DNA synthesis inhibition test), no toxicity being evidenced. MATERIALS AND METHODS: Fifteen male rabbits were used as experimental animals. Rods made from pure lactic acid, pure caprolactone copolymer, and caprolactone-coated lactic acid were placed on both sides of the dorsal muscles, eight implants per rabbit. Rods made from latex and silicone were used as positive and negative controls, respectively. The animals were sacrificed after 1 week, 1 month, or 6 months. Tissue reactions around the implants were analyzed and scored semiquantitatively. RESULTS: Acute tissue reactions attributable to operative trauma were seen in all specimens at 1 week. After 6 months, chronic inflammatory changes and foreign-body reactions were seen only in the positive controls. CONCLUSION: The new caprolactone copolymer material is highly biocompatible.

Absorption↗

Addition of leucine precursors to the diet of leucine-starved mice.

Leucine-starved mice placed on a diet supplemented with the immediate precursor of leucine, alpha-ketoisocaproic acid, regain lost weight. This weight gain is similar to that observed when the leucine-starved mice are provided with leucine in their diet. Mice on a leucine-free diet supplemented with alpha-ketoisovaleric acid, the first compound in the leucine biosynthetic pathway, continued to lose weight as quickly as mice on leucine-deficient diets.

Animals↗

Urinary organic acid excretion during feeding of medium-chain or long-chain triglyceride diets in patients with non-insulin-dependent diabetes mellitus.

Medium-chain triglycerides (MCTs) are absorbed and metabolized differently from long-chain triglycerides (LCTs). Recent data indicate that MCTs may be useful as a dietary substitute in a variety of clinical disorders. The current studies were undertaken to characterize urinary organic acid excretion in patients with non-insulin-dependent diabetes mellitus during 4 d of an LCT or MCT diet. Urinary excretion of the dicarboxylic acids adipic, suberic, and 3-hydroxysebacic and the (omega-1) hydroxylation products 5-hydroxyhexanoic acid and 7-hydroxyoctanoic acid, was increased during MCT feeding as compared with LCT feeding. Urinary suberic and 7-hydroxyoctanoic acid excretions were increased 55- and 30-fold, respectively, during the MCT-substituted diet. Urinary organic acid profiles provide information on the fate of lipids during MCT feeding and may also be useful in assessing complicance during clinical trials employing MCT-substituted diets.

Caproates↗

Protein metabolism in obesity: effects of body fat distribution and hyperinsulinemia on leucine turnover.

To examine whether moderate obesity and differences in body fat distribution are associated with abnormalities of protein metabolism, leucine turnover was measured in three groups of age-matched premenopausal women. Ten upper-body-obese (UB Ob), 10 lower-body-obese (LB Ob), and 10 nonobese (Non Ob) women were studied in an overnight postabsorptive condition (basal) and again during an infusion of low physiologic amounts of insulin (insulin clamp). Results showed that basal leucine carbon flux was greater (P less than 0.05) in UB Ob and LB Ob women than in Non Ob women (2.96 +/- 0.08 vs 3.14 +/- 0.16 vs 2.68 +/- 0.08 mumol.kg lean body mass-1.min-1, respectively; mean +/- SEM). Leucine carbon flux was not suppressed during the insulin-clamp study in UB Ob women but was in the LB Ob and Non Ob women. We conclude that moderate obesity is associated with increased proteolysis and that insulin's antiproteolytic actions are impaired in upper-body obesity. These findings could have implications for future studies of and treatment of obesity.

Adipose Tissue↗

Plasma proline and leucine kinetics: response to 4 wk with proline-free diets in young adults.

The effects of removing proline from the diet on plasma leucine and proline kinetics were investigated. After a 1-wk control period, during which young adult men received a diet containing a complete L-amino acid mixture, seven subjects were given for 4 wk a diet devoid of proline (group 1); six received a diet devoid of proline, arginine, aspartate, glutamate, and serine (group 2); and seven continued with the complete diet (group 3). At the end of the control and 4-wk periods subjects were given a continuous, (3-h fast, 5-h fed) intravenous infusion of L-[1-13C]leucine and L-[5,5-2H]proline. Plasma proline was reduced significantly, especially during the fed state, in groups 1 and 2 after the 4-wk diet periods. Small but statistically significant (P < 0.05) reductions occurred in nonoxidative leucine disappearance and leucine appearance during the fasted state in group 2. Proline fluxes decreased by approximately 50% in fasted and fed states in groups 1 and 2. Mean de novo proline synthesis during the fasted state declined markedly (P < 0.05) after 4 wk in groups 1 and 2.

Adult↗