text:
|Publication number
|Publication type
|Application number
|Publication date
|Mar 11, 2003
|Filing date
|Jun 9, 1998
|Priority date
|Jun 10, 1997
|Fee status
|Publication number
|Edith Mathiowitz, Wendy L. Webber, Christopher G. Thanos
|Original Assignee
|Brown University Research Foundation
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (10), Non-Patent Citations (2), Referenced by (55), Classifications (18), Legal Events (4)
|External Links: USPTO, USPTO Assignment, Espacenet
Priority is claimed to U.S. Provisional application Ser. No. 60/050,946, filed Jun. 10, 1997.
Not applicable.
Effective prophylactic or therapeutic treatment or diagnosis of many illnesses or conditions requires that a relatively constant level of therapeutic or diagnostic agents be administered in vivo. Before the development of time-release delivery systems, prophylactic, therapeutic and diagnostic agents were commonly administered as several doses at regular intervals. However, this mode of administration provides fluctuating levels of these agents.
Sustained release formulations have been prepared to provide a relatively constant release of prophylactic, therapeutic and diagnostic agents. Some of these formulations use biodegradable substances, such as poly(lactide) (PLA) or poly(lactide-co-glycolide) (PLGA) microspheres or films containing the agent to be administered. A relatively steady release of incorporated agents is possible using these microspheres and/or films by taking advantage of the near linear degradation profile of these polymers. However, in some cases, the in vivo release of incorporated agents from biodegradable polymers is initially high or low, and therefore non-uniform throughout the life of the delivery device. Furthermore, some polymers can substantially degrade after a period of hydration, which can substantially limit the effective life of the controlled release devices. Accordingly, several attempts have been made to modulate the controlled release of incorporated agents from biodegradable polymers to provide a higher level of initial medicament release and to provide longer periods of fairly consistent medicament release levels in vivo.
U.S. Pat. No. 4,919,939 to Baker describes polymeric sustained release drug delivery systems for placement in the periodontal pocket. Baker's system incorporates drug-containing microparticles in a carrier medium and is effective for up to 30 days. However, a limitation of the device is that useful diffusion rates through the polymers are generally limited to drugs with molecular weights less than about 300.
U.S. Pat. No. 4,666,897 to Golub et al. teaches sustained release devices made from ethylene vinyl acetate (EVA) for sustained delivery of tetracycline. However, the EVA fibers are non-degradable and therefore must be physically removed after delivery of the therapeutic.
It is therefore an object of this invention to provide compositions for modulated release of an incorporated agent, wherein the composition includes a wide range of biocompatible polymers.
It is another object of this invention to provide biodegradable polymeric compositions for modulated release of an incorporated agent.
Sustained delivery compositions which modulate the release of incorporated prophylactic, therapeutic and/or diagnostic agents, and methods of preparation and use thereof, are disclosed. The compositions include a biocompatible polymeric matrix, a prophylactic, therapeutic, and/or diagnostic agent dispersed within the polymeric matrix, and a monovalent cation component which is separately dispersed within the polymeric matrix. The monovalent cation component modulates the release of the incorporated agent from the polymeric matrix.
The compositions can be prepared by dissolving a biocompatible polymer in a solvent to form a polymer solution, and separately dispersing a monovalent cation and a prophylactic, therapeutic, and/or diagnostic agent within the polymer solution. The polymer solution is then solidified to form a polymeric matrix. At least a significant amount of the monovalent cations is dispersed in the polymeric matrix separately from the incorporated agent. The cation modulates the release of the incorporated agent from the polymeric matrix.
FIGS. 1A and 1B show the cumulative percent (%) of tetracycline released into PBS buffer at 37� C. as a function of time (days). Black squares represent poly (lactide-co-glycolide) (hereinafter "PLGA") solvent cast film loaded with 5% tetracycline (w/w), 0% NaCl. Diamonds represent PLGA film with 1% NaCl and 5% tetracycline. Circles represent PLGA film with 5% NaCl and 5% tetracycline. Triangles represent PLGA film with 25% NaCl and 5% tetracycline. Grey squares represent PLGA film with 25% NaCl, low molecular weight excipient and 5% tetracycline. FIG. 1A shows the first five days of tetracycline release, while the FIG. 1B shows the release over the first 50 days of tetracycline release.
FIG. 2 is a plot of molecular weight (Daltons) as a function of time (days) for polymer films incubated in PBS buffer at 37� C. Grey squares represent a blank PLGA film. Circles represent PLGA film containing 1% NaCl. Dark squares represent PLGA film containing 5% NaCl. Diamonds represent PLGA film containing 25% NaCl. Triangles represent PLGA film containing 25% NaCl and low molecular weight (LMW) polymer excipient.
FIG. 3 is a plot of glass transition temperature (� C.) as a function of time (days) for polymer films incubated in PBS buffer at 37� C. Dark squares represent blank PLGA film. Diamonds represent PLGA film containing 1% NaCl. Circles represent PLGA film containing 5% NaCl. Triangles represent PLGA film containing 25% NaCl. Light squares represent PLGA film containing 25% NaCl and LMW polymer excipient.
FIG. 4 is a plot of percent of original weight of polymer (%) as a function of time (hours) for PLA/PG polymer films incubated in PBS buffer at 37� C.
FIG. 5 is a plot of the original weight of polymer (%) as a function of time for EVA polymer films with 0, 5, and 10% NaCl by weight.
FIG. 6 is a plot of the original weight of polymer (%) as a function of time for polyurethane polymer films with 0, 5, and 10% NaCl by weight.
FIG. 7 is a plot of the original weight of polymer (%) as a function of time for PLA polymer films with 0, 5, and 10% NaCl by weight.
FIG. 8 is a plot of the original weight of polymer (%) as a function of time for PLA/PG polymer films with 0, 5, and 10% NaCl by weight.
FIG. 9 is a plot of the original weight of polymer (%) as a function of time for zein polymer films with 0, 5, and 10% NaCl by weight.
FIG. 10 is a plot of the original weight of polymer (%) as a function of time for EVA polymer films with 0, 5, and 10% pre-dissolved NaCl by weight, and 10, 30, and 50% particulate NaCl by weight.
FIGS. 11A and 11B are graphs showing the cumulative release of RNAse (%) as a function of time for PLGA polymer films loaded with 10% RNAse and 0, 5, and 10% NaCl by weight.
FIG. 12 is a graph showing the percent swelling of PLGA microspheres as a function of time immersed in distilled water for PLGA microspheres loaded with 10% NaCl by weight.
Sustained delivery compositions which modulate the release of incorporated prophylactic, therapeutic and/or diagnostic agents, and methods of preparation and use thereof, are disclosed. The compositions include a biocompatible polymeric matrix; a prophylactic, therapeutic, and/or diagnostic agent dispersed within the polymeric matrix; and a monovalent cation component which is separately dispersed within the polymeric matrix. The monovalent cation component modulates the release of the incorporated agent from the polymeric matrix.
As used herein, modulated release is defined as the change in the release characteristics of an incorporated agent from a biocompatible polymeric matrix containing a dispersed monovalent cation component which is separate from the incorporated agent relative to a polymeric matrix that does not include the monovalent cation component. Release characteristics include the initial release level of the agent, subsequent agent release levels, the amount of agent released, and/or the extent of the release period. The release characteristics can be modified by selecting the type and concentration of the monovalent cation component that is dispersed in the polymeric matrix. In addition, the particle size of dispersed monovalent cation component can selected to modify the release characteristics.
Any biocompatible polymer can be used. As used herein, a polymer or polymeric matrix is biocompatible if the polymer and any degradation products of the polymer, are non-toxic to the recipient and also present no significant deleterious effects on the body of the recipient. The biocompatible polymers can be biodegradable polymers, or non-biodegradable polymers, or copolymers and blends thereof.
As used herein, biodegradable is defined as capable of degrading or eroding in vivo to form smaller chemical species over a period of time between one day and nine months. Degradation can result, for example, by enzymatic, chemical and physical processes. Examples of suitable biocompatible, biodegradable polymers include poly(lactide)s, poly(glycolide)s, poly(lactide-co-glycolide)s, poly(lactic acid)s, poly(glycolic acid)s, poly(lactic acid-co-glycolic acid)s, polyanhydrides, polyorthoesters, polyetheresters, polycaprolactone, polyesteramides, and copolymers and blends thereof.
Suitable non-biodegradable polymers include polyacrylates, polymers of ethylene-vinyl acetates and other acyl substituted cellulose acetates, non-degradable polyurethanes, polystyrenes, polyvinyl chloride, polyvinyl fluoride, poly(vinyl imidazole), chlorosulphonate polyolefins, polyethylene oxide, blends and copolymers thereof.
The end-groups of the polymers can be blocked or unblocked. Suitable blocking groups include alkyl groups. Acceptable molecular weights for the biocompatible polymers can be determined by a person of ordinary skill in the art of taking into consideration factors such as the desired polymer degradation rate, physical properties such as mechanical strength, and rate of dissolution of polymer in solvent. Typically, an acceptable range of molecular weight is between about 2,000 and 2,000,000 Daltons (Da). In a preferred embodiment, the polymer is a biodegradable polymer or copolymer. In a more preferred embodiment, the polymer is a poly(lactide-co-glycolide) (PLGA) with a lactide:glycolide ratio of about 1:1 and a molecular weight between about 5,000 Da and 70,000 Da. In a more preferred embodiment, the molecular weight of the PLGA is between about 5,000 Da and 42,000 Da.
Agent that can be incorporated into the polymeric matrix include therapeutic, prophylactic, and diagnostic agents. Examples of suitable therapeutic and/or prophylactic agents include proteins, such as hormones, antigens, and growth factors; nucleic acids, such as antisense molecules; and small molecules, such as antibiotics, steroids, decongestants, neuroactive agents, anesthetics and sedatives. Examples of suitable diagnostic agents include radioactive isotopes and radiopaque agents. The polymeric matrices can include more than one incorporated agent.
A therapeutically, prophylactically, or diagnostically effective amount of the agents are incorporated into the polymeric matrices. An effective amount of these agents can be readily determined by a person of ordinary skill in the art taking into consideration factors such as body weight; age; physical condition; therapeutic, prophylactic, or diagnostic goal desired; type of agent used; type of polymer used; initial burst and subsequent release levels desired; and desired release rate. Typically, the polymeric matrices will include between about 0.01% (w/w) and 80% (w/w) of incorporated agent.
The incorporated agent may be in the form of particles, for example, crystalline particles, non-crystalline particles, freeze dried particles, and lyophilized particles. Particles preferably are less than about 20 μm in size, and more preferably less than about 5 μm. The particles also may include a stabilizing agent and/or other excipient.
A monovalent cation component, as defined herein, is a component containing at least one kind of monovalent cation (having a valence of +1) in a non-dissociated state, a dissociated state, or a combination of non-dissociated and dissociated states. Suitable monovalent cation components include, for instance, substances such as metal salts. It is to be understood that an anion is associated with the cation.
The particle size of the monovalent cation component preferably is less than about 50 μm and more preferably less than about 20 μm. Even more preferably, the particle size of the monovalent cation component is less than about 5 μm. Generally, smaller particle sizes provide better modulation of release of the incorporated therapeutic, diagnostic, or prophylactic agent, than larger ones. If very large particles, such as those 100 micron and larger in size, are used, then the swelling effect of the polymer is diminished and the particles only leach out. However, when the particle size is smaller, the entire polymer system swells. Swelling is the mechanism by which modulation occurs.
A suitable concentration of a monovalent cation component is dispersed within the polymer matrix. Suitable concentrations are those which modulate the release of incorporated agents from the polymeric matrix. The optimum concentration depends upon the polymer, the monovalent cation component and the biologically active agent utilized. Preferably, the concentration of the monovalent cation component is between about 1% and 30% (w/w) of the total weight of the polymer matrix, including the incorporated agent. More preferably, the concentration of the monovalent cation component is between about 1% and 20% (w/w) of the total weight of the polymer matrix. Monovalent cation component concentrations greater than about 50% (w/w) are not preferred, since higher concentrations generally lead to leaching out of the monovalent cation component without polymer swelling.
In one embodiment, the monovalent cation component is substantially soluble in aqueous solutions, such as PBS, HEPES, or alimentary tract fluids. Examples of suitable soluble monovalent cation components include NaCl, NaF, KCl, KF and combinations thereof.
The polymeric matrix can be prepared by dissolving a suitable polymer in a solvent to form a polymer solution, adding an aqueous solution or dispersion of the monovalent cation composition to the polymer solution, and adding a solution or dispersion of the agent to be incorporated. Addition of the monovalent cation component can be completed before addition of the therapeutic, diagnostic, or prophylactic agent. For example, the polymer solution and the salt solution or particles can be mixed by sonication or agitation, while the agent is incorporated later in the process of forming the polymeric matrix.
In those embodiments in which the polymer is insoluble in aqueous solutions and soluble in organic solvents that are immiscible with water, an emulsion can be formed. Emulsions can be formed, for example, by sonicating, agitating, mixing, or homogenizing these solutions.
In a preferred embodiment, the method includes forming a modulated release polymeric matrix as a thin film. Biodegradable polymers are preferred; PLGA is more preferred. A suitable monovalent cation component is dissolved in distilled water and sonicated into the polymer solution along with a biologically active agent also dissolved in distilled water. A thin film is then solvent cast from the polymer solution and left to dry overnight. The film is then subjected to high vacuum for a period of 4-6 hours to extract any residual solvent. The formation of polymeric matrix films is further described in Example 1.
In another embodiment, the method includes forming a modulated release system via the spray drying process. Alternately, the method includes forming modulated release polymer microspheres via the solvent removal process. Either method forms microspheres, or microparticles, encapsulating the monovalent cation component and biologically active agent within the system.
The type of solvent used to dissolve the polymer will depend on the type of polymer. Suitable solvents for dissolving the polymers include polar organic solvents such as methylene chloride, chloroform, acetone, ethyl acetate, tetrahydrofuran, dimethyl sulfoxide, and hexafluoroisopropanol.
In one embodiment, particles of at least one monovalent cation component are pre-dissolved in distilled water and then dispersed within the polymer solution. At least one biologically active agent is added to the polymer solution separately from the addition of the monovalent cation component solution. The biologically active agent can also be dissolved in distilled water, thereby adding to the polymer and cation component emulsion. The monovalent cation component and the biologically active agent can be added to the polymer solution sequentially, in reverse order, intermittently, or through separate, concurrent additions. A biologically active agent can be suspended in a solution of a monovalent cation component in a solvent before dissolving the polymer in the solvent.
In another embodiment, the monovalent cation component is incorporated into the polymeric matrix after the matrix has been formed and has already incorporated the agent. In an alternate embodiment, the protein or active drug added to the polymer solution can be mixed with an excipient, such as at least one stabilizing agent, as is known in the art.
The amount of a biologically active agent added to the polymer solution can be determined empirically by comparative in vitro tests of polymeric matrices containing different concentrations of at least one monovalent cation component and of at least one biologically active agent. The amount used will vary depending upon the particular agent, the desired effect of the agent at the planned release levels, and the time span over which the agent will be released.
Several types of delivery devices, such as thin films, pellets, cylinders, discs, and microparticles can be prepared from the polymeric matrix, using methods well known to those of skill in the art. As used herein, microparticles are particles having a diameter of less than about one millimeter that include an incorporated agent. The microparticles can have a spherical, non-spherical, or irregular shape. Preferably, the microparticles are spherical.
Microspheres formed by the solvent evaporation process are not contemplated to be within the microspheres disclosed herein, unless they were left for a very short time to harden. Otherwise, the monovalent cation component would leach out of the system during the fabrication of the system.
In another embodiment, the method includes forming a modulated release polymeric matrix as a cylinder or any other shape. A polymer solution and monovalent cation component, in dissolved form, are mixed, for example by sonication, until a fine emulsion is produced. The polymer solution is subsequently cast into a mold of the desired shape. The solvent is then removed by means known in the art until a cylinder or other form, with a constant dry weight, is obtained.
Several other methods of using the polymeric compositions include modulating the physical properties of a method for modifying the water absorption, or hydration capacity, without significantly affecting the rate of polymer degradation. The method includes forming a solution of a polymer and then dissolving a monovalent cation component and sonicating it into the polymer solution. The polymer solution is then solidified to form a polymer matrix wherein the monovalent cation component is dispersed therein. See Example 6 below for a further description of this method of enhancing initial polymer hydration.
The composition can be administered to a human, or other animal, for example, by injection and/or implantation subcutaneously, intramuscularly, intraperitoneally, intradermally, intravenously, intraarterially or intrathecally; by administration to mucosal membranes, such as intranasally or by means of a suppository, or by in situ delivery to provide the desired dosage of a biologically active agent based on the known parameters for treatment of the various medical conditions with the agent.
The compositions and methods of preparation and use thereof described herein are further described by the following non-limiting examples.
Blocked-PLA/PG (50:50) with a molecular weight of 35,000 Da (Boehringer Ingelheim, Resomer RG503, lot #223808) was used for all studies. The films were produced by a film casting technique. The polymer was thoroughly dissolved in methylene chloride (10% w/v) at room temperature.
Films were prepared using water soluble salts containing monovalent cations. The salts were incorporated in the polymer as an emulsion via probe sonication after predissolving the salt in distilled water. The fabrication procedure is described below.
For each set of control films, one gram of PLA/PG polymer (Boehringer Ingelheim, Resomer RG503, lot #223808, MW=34,000 Da) was dissolved in 10 ml methylene chloride (Fisher Scientific) in a glass scintillation vial. A loading of 0%, 1%, 5% or 25% (w/w) sodium chloride (Sigma, lot 105H02501) pre-dissolved in 200 microliters of distilled water was also added to each vial. For each film composition, a set of tetracycline-loaded films was also made. 5% (w/w) tetracycline, also pre-dissolved in 100 ml of distilled water, was used. The contents of the vials were then sonicated for 60 seconds each and cast into ten circular glass rings on Teflon sheets. Films were air-dried at room temperature for 24 hours before being subjected to high-vacuum for an additional 4 hours to extract any residual solvent.
The purpose of this experiment was to test the effect of salt incorporation on drug release from the film samples. Films were immersed in 1.0 M solution of PBS buffer (FTA hemagglutination buffer) at 37� C. for the duration of the study. At regular intervals, the buffer was extracted from the vials and replaced with fresh buffer. The buffer samples were analyzed spectrophotometrically at 354 nm for tetracycline quantification using a Beckman DU-65 spectrophotometer. FIGS. 1A and 1B shows the tetracycline release curves for the various salt loaded systems over the short term (0-5 days) and the long term (0-50 days), respectively.
It is evident from these curves that the soluble salts have a significant impact upon drug release from the polymer films. The control films loaded with 5% tetracycline seemed to follow a typical release pattern for PLA/PG, with an initial release of drug (probably due to diffusion), followed by a plateau phase, another sharp increase in release (possibly due to polymer degradation), and then a second plateau phase. The incorporation of monovalent salts appears to beneficially control the initial burst of drug which occurred in the first few days of release in the control films.
The 1% NaCl system achieved slow, sustained release of tetracycline from the drug-loaded films for periods exceeding six weeks. This system released 12.39% of the drug per day for the first week, decreased to a release of 4.67% of the drug over the next two weeks, and then leveled off at a release rate of 0.01% per day for the last 3 weeks.
The 5% NaCl system also released drug over a six week period, although more of the drug is released in the initial weeks than in the 1% NaCl system or control system. Here, 23.84% of the drug per day came out of the film in the first two days. The rate of drug release then dropped to 0.91% per day from week 1 to week 3. Next, the drug release leveled off at a rate of 0.005% drug release per day from week 6 to week 23 before rising to a rate of 0.05% release per day for the final 3 weeks of the study. Even in this case, the three phase release, as seen in the control samples (0% NaCl), was not observed.
The use of 25% salt resulted in an increased release rate, with the majority of the drug diffusing from the film within the first three weeks. The rate of release was measured at 58.27% per day for the first day, decreased to 0.76% per day for the next two weeks, and leveled off at a release rate of 0.003% drug release per day from week 6 to week 20 of the study.
The 25% salt system containing low molecular weight excipient (PLA/PG MW=5,000 Da) resulted in the greatest amount of release, with most of the drug being expelled in one week. 50.85% of the drug load was released per day in the initial 24 hours of the study, followed by a rate of 8.11% release per day for the next two days, and leveling off early at a release rate of 0.0007% per day for weeks 2-15.
The results clearly show that the salt significantly modulates the drug release from the PLA/PG films. Sodium chloride is very water soluble, so one would assume that most of the salt would come out in the first few days. However, this assumption proved inaccurate for the 1% and 5% salt-loaded systems. Therefore, the following experiments were undertaken to further characterize the effects of salt on the release rate of tetracycline.
Surface morphology of the films was analyzed over time for evidence of pore formation and subsequent film degradation. Scanning electron microscopy (SEM) was performed using a Hitachi S-2700 scanning electron microscope on film surfaces sampled at day 0, 1, 3, 7, 21 and 42 days in buffer. Film samples were frozen and lyophilized, mounted on metal stubs, and sputter-coated with gold-palladium (Polaron Instrument E5100) prior to viewing.
Films with different salt loadings and from various representative time points were examined under scanning electron microscope for changes in surface morphology. The films degraded over time, as indicated by the formation of pores in the polymer matrix. All film surfaces appear smooth and homogeneous at time zero. However, after being immersed in PBS buffer, the pores form on the film surfaces, indicating degradation and/or swelling. At 0, 1, and 3 days, no change in the appearance of the surface of the control films is evident. At 7 and 21 days, the surface of the control film was still relatively smooth and pore-free, although a few minor blemishes appeared. It was not until 42 days that the control film developed actual pores 5-10 μm in diameter, which appeared rough and in a beehive-like configuration.
In the 1% NaCl-loaded system, small irregularities in the smooth surface appeared at days 1 and 3. By 7 days, the surface was covered with pinholes 1-5 μm in size. These pores increased to 5 μm in size by 21 days and resulted in a rough and rugged, degraded appearance by 42 days.
Similarly, the 5% NaCl-loaded system remained pore-free at 1 day, but had small pinholes 1-5 μm in size in its smooth surface by 3 days. At 7 and 21 days, the film was homogeneously covered with distinct pores 5-10 μm in size. After 42 days in buffer, however, the film surface degraded away completely, leaving a rough, irregular surface.
The 25% NaCl system formed pinhole pores 1-5 μm in diameter even prior to submersion in buffer. The pores remained this size for days 1 and 3. However, these pores enlarged significantly, resulting in pores up to 20 microns in diameter at 7 days. At this point, the polymer matrix looked much like a beehive. By 42 days, the surface was again rough and irregular in appearance, as if the structural integrity of the 'beehive' had broken down.
Lastly, the high and low molecular weight blend system with 25% NaCl had small pores over the entire surface at days 0, 1 and 3 days. By 7 days, the film was covered with 30 micron pores. This system degraded too rapidly after this time for further measurements to be taken accurately.
Gel permeation chromatography (GPC) on a Perkin Elmer liquid chromatography system was used to determine molecular weights and molecular weight distributions via size-exclusion chromatography. The Perkin Elmer system consisted of an Isocratic LC pump model 250, LC-30 RI detector, LC column oven model 101, and 900 series computer interface box. Samples were lyophilized, dissolved in HPLC-grade chloroform and filtered with a 0.2 micron syringe filter before injecting to remove any insoluble particulates. The samples were then passed through a PL gel 5 micron mixed column and a 5 micron/50 angstrom column at a flow rate of 1.0 ml/min and a temperature of 40� C. All samples were compared to polystyrene standards (Polysciences) ranging in molecular weight from 1000 to 1,860,000 Da. Turbochrom and TC*SEC software programs from Perkin Elmer were utilized for data analysis. Polymer films were sampled at each of the specified time points, including 0, 1, 3, 7, 21 and 42 days.
Tables 1 and 2 indicate degradation of the films as monitored by GPC. The molecular weights decreased as time in buffer increased. Despite the variation in salt loading, the basic trend in molecular weight decrease is similar in most salt systems, except for the polymer blend system. Soluble salts added to the system have very little effect on trends of molecular weight decrease. The blend system, however, utilized a low molecular weight excipient to hasten degradation, speed up release, and appeared to degrade the fastest, as the molecular weight was no longer measurable after one week in buffer. The 5% and 25% NaCl systems, on the other hand, seemed to decrease in molecular weight more slowly than the other systems. FIG. 2 is a graph showing the change in molecular weight over time for the five different systems, with 0, 1, 5, and 25% NaCl, and also 25% NaCl with a low molecular weight excipient. It is evident from the data that the tetracycline-loaded films follow the same pattern as the control films.
GPC Molecular Weights of Salt-loaded
PLA/PG Control Films
# Days in
25% NaCl +
0% NaCl
1% NaCl
5% NaCl
25% NaCl
*There was not enough sample remaining to properly analyze.
GPC Molecular Weights of Salt and
Tetracycline-loaded PLA/PG Films
# Days in
25% NaCl +
0% NaCl
1% NaCl
5% NaCl
25% NaCl
*There was not enough sample remaining to properly analyze.
Differential scanning calorimetry (DSC) was used to determine the glass transition temperature (Tg) of the polymer films. All film samples were lyophilized for 24 hours before DSC was performed on a Perkin Elmer Model DSC 7 connected to a controller model TAC 7/DX (Perkin Elmer). Samples were run for each of the following time points; 0, 1, 3, 7, 21, and 42 days after immersion in PBS buffer. Samples were heated from −20 to 100� C. at a rate of 10� C./min, cooled back down to −20� C. at the same rate, and finally heated to 100� C., again at 10� C./min. Glass transition temperatures were calculated from the second heating ramp and examined for shifting trends which would indicate the occurrence of film degradation. Analysis was performed using the Perkin Elmer Thermal Analysis software.
The glass transition temperature of the control films increased over the first few days, then showed a dramatic decrease over the next few weeks. At day 7, the Tg was below the glass transition of PLA/PG. This correlated with the time at which the molecular weight decreased significantly. However, pores were not visible on the control films under SEM at the 21 day point, which could be explained by the fact that the system collapsed due to the low Tg.
Samples of 1%, 5%, and 25% salt-loaded PLA/PG films, as well as the 25% salt-loaded high and low molecular weight blend, were also lyophilized and subjected to differential scanning calorimetry (DSC). Thermograms were analyzed for changes in glass transition temperature, which would indicate film degradation. In Tables 3 and 4 below, it is evident that very little change occurred in the glass transition temperature in the 1%, 5% and 25% salt systems until the 21 day time point, when the Tg fell below the glass transition of PLA/PG. This is also the time at which the molecular weight of these films, as seen by GPC, dropped dramatically and pores 10-50 microns in diameter were visible under SEM.
Although the glass transition of the 25% salt system began dropping at the same time, it was seen to make a more dramatic drop in Tg at 42 days, at which point the molecular weight of the system dropped down to 965-997. Using scanning electron microscopy, it appeared that by 21 days the film was composed primarily of large voids and had likely lost most of its material. At that point, the Tg had dropped below the known glass transition temperature for PLA/PG and descended to a low 3.7� C. by 42 days. The 25% NaCl system with low molecular weight excipient noticeably dropped in Tg by one week, then rapidly degraded such that no further thermograms could be taken. FIG. 3 is a graph illustrating the change in glass transition over time for the various systems. The tetracycline did not seem to impact the system, as the DSC results followed the same trend in both systems.
DSC Glass Transition Temperatures (� C.)
of Salt-loaded PLA/PG Control Films
# Days in
25% NaCl +
0% NaCl
1% NaCl
5% NaCl
25% NaCl
*These samples were degraded too much for accurate thermograms to be taken.
DSC Glass Transition Temperatures (� C.) of Salt
and Tetracycline-loaded PLA/PG Films
# Days in
25% NaCl +
0% NaCl
1% NaCl
5% NaCl
25% NaCl
*These samples were degraded too much for accurate thermograms to be taken.
Poly (lactide-co-glycolide) films (or PLA/PG films) containing various concentrations of salt, including 0%, 1%, 5%, 10%, 20% and 25%, were preweighed and left to incubate in PBS buffer. At regular intervals, the samples were removed from buffer and weighed. This experiment was performed to investigate the effect of the incorporation of soluble salts on water uptake and swelling of the polymer film. FIG. 4 shows the water uptake of the PLA/PG films with different salt concentrations. Blank films had a small amount of swelling at two hours, and then rapidly returned to their original weight for up to 50 hours. Salt loaded films, on the other hand, demonstrated increased swelling of over 200%, depending on the loading of salt in the system. "A" and "B" indicate two different experiments, and both demonstrate in FIG. 4 that only the salt-loaded films swell appreciably, while the blank films do not.
Polymer films containing 0%, 5%, and 10% concentrations of salt were weighed and left to incubate in PBS buffer. At regular intervals, the samples were removed from buffer, air dried, and weighed. Polymers used in this experiment were ethylene vinyl acetate (MW 235 kDa), Cardiothane (a medical grade polyurethane), poly(lactic acid) (MW 300 kDa), poly(lactide-co-glycolide) (MW 34 kDa), and zein. FIGS. 5-9 show the increased swelling of salt loaded polymer films prepared from EVA (FIG. 5), polyurethane (FIG. 6), PLA (FIG. 7), PLA/PG (FIG. 8), and zein (FIG. 9). The results showed that a significantly greater amount of swelling occurred in rubbery or amorphous polymers, such as ethylene vinyl acetate, as compared to semi-crystalline polymers, such as PLA/PG. For example, PLA did not show significant swelling, possibly indicating that the salts affect amorphous polymeric materials more easily than crystalline polymers.
Another experiment was performed to determine the effect that salt particle size has in the role of water uptake. Ethylene vinyl acetate (EVA) films were made containing 0%, 5%, and 10% loadings of soluble salt that had been pre-dissolved in 200 ml of distilled water and compared to EVA films made containing 10%, 30%, and 50% loadings of soluble salt that had been left as particulates 212 μm in size. FIG. 10 illustrates the difference in the extent of water uptake that results from varying the salt particle size in a particular polymer. Virtually no swelling occurred in systems loaded with large particles, since they dissolved quickly and leached out of the system, in contrast to small particles that cause the system to swell.
X-ray diffraction techniques, performed on a Siemens Diffraktometer D5000, were used to measure film degradation analysis. Samples that had been in buffer for 0, 1, 3, and 7 days were subjected to x-ray and analyzed using DiffracAt computer software for decreasing or disappearing representative crystalline or salt peaks, as well as growth changes of other peaks or amorphous areas. The X-ray diffractometer was also used as a means of determining how long salt remained a part of the system. X-ray diffraction studies on a number of polymers incorporating monovalent salts demonstrated that at least a percentage of the salt remains in the system after more than one week in buffer.
By X-ray diffraction, the salt loading can be measured by looking for a characteristic peak for NaCl in the 32 range on the two-theta axis. The height of this peak increases in size as the loading of salt increases from 5% to 25%, indicating an increase in crystallinity. Similarly, pure PLA/PG polymer has its most pronounced peak appearing at 30, with smaller peaks at 25, 29, 36, 40, 43, 48 and 49. Evidence of salt leaching and decrease in crystallinity was seen as the salt peak for both the 5% and the 25% systems then decreased over time as the salt dissolved and washed out of the system. The decrease in the peak appearing around 32 occurred at day 1, which indicated that salt dissolution begins upon introduction into the buffer solution. The occurrences were most pronounced in the 25% salt system. However, the salt peak in both systems was still evident after one week in buffer, showing that not all of the salt leaches out of the system immediately. This data suggests that the salt is stabilizing the system in some manner, leading to the controlled release.
The purpose of this experiment was to test the effect of salt incorporation on release of proteins (or larger molecular weight species) from film samples. PLGA 50:50 films (Boehringer Ingelheim, Resomer RG 503, lot #223808, MW=34,000 Da) containing 10% RNAse (w/w) and 0%, 5%, or 10% loadings (w/w) of NaCl were made via the solvent casting technique. Films were immersed in 1.0 M solution of PBS buffer (FTA hemagglutination buffer) at 37� C. for the duration of the study. At regular intervals, the buffer was extracted from vials and replaced with fresh buffer. The buffer samples were analyzed using the BCA Protein Assay and spectrophotometric measurement at 562 nm to quantify the amount of RNAse released from the systems over time. FIGS. 11A and 11B show the release curves for the various salt loaded systems over the short term (0-5 days) and the long term (0-40 days), respectively.
The 0% NaCl films (controls) appear to follow a classical release pattern from PLGA, in which there is an initial release of protein, followed by a lag phase, which is followed by a second release beginning around 310 hours, which is likely due to degradation of the polymer. Again, additions of the salts altered this typical triphasic behavior, resulting in more consistent release of protein. The salt also provided for an overall greater amount of protein released. The control film released only about 18% of its protein load in 500 hours, while the 5% and 10% NaCl loaded films released about 55% and 58% of their protein loads in 500 hours, respectively. The results, as shown in FIGS. 11A and 11B, show that addition of the monovalent salt significantly modulates the release of proteins from the PLGA films, despite the much larger molecular weight of the species to be released.
Microspheres were made from low molecular weight PLGA (MW=5,800 Da) using a modified solvent removal technique. Particles of sodium chloride (NaCl) were created by spray-drying and crushing with a mortar and pestle, to yield particles less than 5 μm in size. Next, 5 ml of a 10% solution of the PLGA in ethyl acetate and 10% (w/w) NaCl was placed in a separatory funnel with a probe sonicator fitted concentrically within the funnel. The valve on the funnel was adjusted such that pulsed sonication resulted in distinct drops every 4 seconds. Sonication amplitude was set at 20% and was useful in maintaining a homogeneous suspension of salt and polymer. The solution was dropped into a 400 ml glass beaker filled with 100 ml of heavy mineral oil spinning at 1800 rpm with the impeller slightly off-center. After 30 minutes, the speed was slowed to 1400 rpm and 100 ml petroleum ether was slowly dripped in for 4 hours. The resulting microspheres were removed by vacuum filtration.
One microsphere was isolated under a stereoscope using forceps and was placed on a slide with a 1 mm graded line. This slide was immersed in a petri dish filled with 20 ml distilled water and was observed using light microscopy and an attached camera. The experiment was done at room temperature, although the light source on the microscope was turned on during picture-taking which could have generated some heat. Pictures were taken at 0.1, 0.25, 1, 2, 4, 5, 8, 12, and 26 hours. The negatives of these pictures were scanned and measured using NIH imaging software. The percent swelling is defined as total area at a certain time point divided by the initial area of the microsphere at t=0. The results are shown in FIG. 12, which indicates significant swelling, especially in the first 5 hours of the experiment.
Modifications and variations of the polymeric matrices and methods of preparation and use thereof will be obvious to those skilled in the art from the foregoing detailed description. Such modifications and variations are intended to come within the scope of the appended claims.
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|U.S. Classification
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|International Classification
|Cooperative Classification
|European Classification
|Jul 20, 1998
Owner name: BROWN UNIVERSITY RESEARCH FOUNDATION, RHODE ISLAND
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:MATHIOWITZ, EDITH;WEBBER, WENDY L.;THANOS, CHRISTOPHER G.;REEL/FRAME:009441/0881;SIGNING DATES FROM 19980619 TO 19980701
|Sep 5, 2006
|Fee payment
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Monfragüe, Extremadura's only national park, lies at the heart of the province of Cáceres, close to the confluence of the rivers Tajo and Tiétar, which carve out deep gorges as they run through a series of low mountain ranges. The resulting rocky cliffs offer an ideal location for birds to breed.
- Where: Cáceres
- Ideal for viewing: peregrine falcon, Spanish imperial eagle, Bonelli's eagle, river otter, Eurasian black vulture
- Excellent for: Walking safaris, Vehicle safaris, Birdwatching

subset: commoncrawl


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Warm-up: 500m Row, 50 DU, Hip Joint Mobility
Strength: 4Sets of 7FS/13BS @65% of your FS 1RM
(Complete 7 FS, rack the bar, no rest, complete 13BS @same weight immediately. If you don't know your FS 1RM, find out today!!)
1min ME HSPU
2min ME KB Snatch+OHS (alt every 5)
1min ME T2B
1min Rest
Mobility Video: Super Couch Stretch https://www.youtube.com/watch?v=5EiUquYdyPU


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When it comes to recruiting new talent, firms are looking outside traditional recruiting routes and beginning to embrace social media as a huge recruiting tool. Twitter is quickly becoming the top social media channel for recruiting, and for good reason. AkkenCloud has created a graphic entitled "How to Use Twitter for Recruiting", explaining why the little blue bird holds huge promise for recruiters and jobseekers.
For absolutely no cost, recruiters are able to tap into huge candidate pool, where they can share job postings, connect with applicants, do brand promotion, and even hold twitter chats. For the 56% of jobseekers that use Twitter to scope out new positions, Twitter can be a huge resource of information and a way to connect directly with the company.
With such a fantastic way of connecting jobseekers with employment, it's no wonder that the social media site is such a valuable tool.
Via AkkenCloudBack to Recruitment blogs


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2014 Bold Pitcher League Leaders
Yesterday, I went beyond the obligatory 10 bold predictions and shared my bold hitter league leaders in each of the five standard fantasy categories. So today, I take on the pitchers. Although still difficult to hit on any, the pitchers are a bit easier to get right than the hitters for two reasons. One, there are two ratio categories, rather than one, so playing time expectations are less important. And two, there's a larger luck component involved in pitching which makes the elite guys less of a lock to lead the league in any specific category.
In 2012, I hit on one of my 10 picks. Last year, I was demoted back to the minors after putting up an 0-fer. Let's hope that lit a fire under my butt and results in a breakout predictions year.
American League
ERA – Corey Kluber
I debated whether Alex Cobb and Danny Salazar would be considered bold and decided that they would not be. So Kluber is the pick. Kluber finished seventh in the AL last year in SIERA among pitchers who threw at least 140 innings. I'm a proud member of the Society and believe that his breakout was legit. The only thing he needs is some better defensive support to bring that BABIP down and his HR/FB rate to drop toward the league average.
WHIP – R.A. Dickey
He was one of my Pod's Picks for a reason. A healthier Dickey with better knuckleball velocity enjoyed a skills rebound during the second half of the season. He possesses excellent control and knuckleballers do have the ability to prevent hits on balls in play. Those ingredients mix to create a recipe that results in a pretty darn good WHIP.
Strikeouts – Scott Kazmir
Blah, unless Yu Darvish gets hurt, any other name represents pretty poor odds. Kazmir's career high innings total is just 206 and he has already had a triceps issue during spring training. But his strikeout ability is back now with a return of his velocity and he punched out over a batter per inning last season. Obviously, innings will be key here. Even with 200 innings, he has little chance of taking the strikeout title, so this is more of an "I like Scott Kazmir" thing than really believing he has a chance to lead the league in Ks.
Wins – Sonny Gray
This was a tough one. I think Gray will probably be a bit overvalued in most leagues, but he owns an intriguing skill set. It's one that includes the holy skills trifecta. He threw a total of 182 innings last year, so it wouldn't be crazy to think he can go 200 this season. The Athletics continue to run out a solid and underrated offense, so run support shouldn't be a major problem.
Saves – Nate Jones
There aren't many closers that would truly be considered a bold choice as we are all well aware of how fluky the saves stat is. The White Sox have not officially named a closer yet, but you figure it's going to be Jones who gets the first shot. He throws really hard and induces a healthy rate of ground balls. With no other standout candidates in the Sox bullpen knocking on the door, there's no reason to believe that Jones shouldn't hold the job all year, and perform quite well.
National League
ERA – Alex Wood
Like with AL strikeouts, any pick not named Clayton Kershaw is highly unlikely. This may be cheating a bit because Wood is almost a lock to spend some time in the bullpen, which should benefit his ERA. As I mentioned in my 10 bold predictions article, I love his ability to generate grounders while also inducing swings and misses and displaying good control. He'll need to pitch at least 162 innings to actually qualify for the ERA title, which may be beyond the upper limit of what the Braves plan to do here.
WHIP – Doug Fister
Assuming his elbow issue is nothing, Fister should really benefit from the move to the National League. His strikeout rate should get a boost and he should receive better defensive support, reducing his hits allowed total. He has posted a WHIP as low as 1.06 before, so it's not outlandish to believe he is capable of producing a WHIP low enough to compete with the Kershaws and Cliff Lees of the world.
Strikeouts – Jose Fernandez
Bold? Not bold? He finished 14th in the league in strikeouts last year, so I'm voting bold. I am projecting him to post the second highest K/9 in the league, so it's going to be an innings thing. He threw 172 innings last year, so can he get to 200-210 this time around?
Wins – Matt Garza
A (hopefully) full season of Ryan Braun, along with new left fielder Khris Davis, the Brewers sport one of the best offense in the National League. Garza has thrown 200 innings twice over his career, so if healthy, we know he has the stamina to do so.
Saves – Steve Cishek
Why not? He has excellent skills, combining a strong ground ball rate with a high strikeout rate and improved control. Despite featuring a fastball/slider combo, he was still excellent versus left-handed batters, limiting them to a .288 wOBA and posting a 3.21 xFIP last season. He doesn't get the fanfare, but he's as good as any other closer beyond the top tier.
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Bus to Malaysia – Buy Online And Getting Cost-Effective Travel
The Internet exists at all corners of the world. No wonder, the number of business that has been integrated online is getting higher than ever. They really know that The Internet is the key to higher profit. It is easy to attract consumers since they offer the flexibility, ease, and convenience that cannot be found in a conventional transaction. Now, people do not have to stand in a long queue to get the wanted goods. Now, you can get the tickets bus to Malaysia through online booking. There are many benefits of purchasing the ticket online.
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You might a little bit skeptical about purchasing tickets bus to Malaysia online. Is it possible? Do you have to believe it? Well, of course, there are numbers of benefits you can get from this transaction. First, you do not have to go anywhere to get the tickets. Literally, you only have to sit down, find the most comfortable position, and use your gadget. You do not have to go to the ticket shelter anymore. It saves your time. In addition, you can buy the ticket anytime you want. It can be in the lunch time, on the way back home, or while you are doing your house chores. This is the flexibility offered by online bus booking.
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This bed and breakfast type establishment is a wonderful way to spend a weekend or a week.
Ocean front, beautiful views, rocking chairs on the front porch. Complementary continental breakfast, afternoon tea and cookies, and port and wine served with cheese in the early evening. And coffee or tea at any hour of the day or night in the lobby. The room we stayed in did not face the ocean, but it was lovely, even though it was smallish. There were complementary white his and hers robes, which was a nice touch. Staff could not have been more accomodating. Our towel rack broke and we mentioned it to the maid, within minutes a handyman came to the room and did the repair. I recommend this place to anyone who wants a relaxing getaway. We will definitely stay here again.
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- Also Known As:
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Traditional wear isn't just for weddings and family events – you can incorporate elements of it into your daily ensembles
Mastering the art of business casual
While the corporate dress code has become a bit more lax over the years, this doesn't mean you shouldn't try to look your best with the least effort possible. Here are four items to help you do that
Three ways to do utility wear
Utility wear was a way to survive for most during the 1940s when materials such as wool and leather were in short supply
Avoid these fashion mistakes at work
Despite all the pressure to look good at work, trying too hard is not the way to go. Here are the fashion mistakes gents need to stop making at the office
Lubabalo Kibido: the man behind Gemini Handcrafts
We chat to Lubabalo Kibido, founder and creative director of Gemini Handcrafts, an upcoming luxury leather brand
The details of denim
There are a wide range of denims out there, each with a particular look and feel. Knowing what's what will help you make a better choice for your body, personality and lifestyle
Trends from SA Menswear Week
We bring you the key trends and themes for summer straight off the runway at the fourth SA Menswear Week (SAMW) in Cape Town
Essential spring items to invest in during winter
There's no harm in shopping for spring items slap bang in the middle of winter. Someone once described this as looking ahead to "a brighter future"
iFani's style (r)evolution: an image upgrade of note
If you haven't noticed yet, rapper iFani has upped his dressing game and is now the epitome of a refined gentleman
The dos and don'ts of winter fashion
Winter is not the season to be clumsy, but the season to be your most stylish. Here are a few dos and don'ts to have you well on your way to being a classic style winter gentlemen
SA's most stylish footballers
Soccer players can also be style stars off the field, and here are a few of the guys who take their style very seriously
Style tips from SA's women of fashion
We asked several female designers and stylists for their top style advice for men… read and learn!
How to look like a millionaire
Before you make a million bucks, you have to look like a millionaire, so here are a few tips to get you on your way
Your guide to buying a top quality watch
Buying a watch is not as simple as buying a pair of socks or a T-shirt, so here's some essential info to help you ensure you're making a wise investment


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Through his first five games with the NHL's Colorado Avalanche, forward Gabriel Bourque has failed to record a point.
As a result, he's headed down to the AHL.
The Avalanche confirmed on Monday morning, via the team's official Twitter account, that Bourque is headed to the San Antonio Rampage after serving as an extra forward or bottom six roster member through the start of Colorado's season.
Drafted 132nd overall by the Nashville Predators in 2009, the 26-year-old forward skated for the Baie-Comeau Drakkar and the Moncton Wildcats of the QMJHL before going pro in Nashville's system in 2010.
While with Nashville – the only NHL club Bourque has skated with before joining Colorado this summer – the Rimouski, Quebec native skated out in 242 career NHL games, putting up 31 goals and 78 points in the process.
Last season, he only saw 22 games' worth of action in Nashville, spending time with the AHL's Milwaukee Admirals as well before ultimately being let go by the Central Division club over the summer. He signed on as a free agent with Colorado this fall, who inked him to a one year deal worth $800,000 at the highest level.
For Colorado, things have turned around this year after watching head coach Patrick Roy depart of his own volition over the summer. As a result, they're likely feeling out what parts of their roster are the best for a winning season, and which parts are extraneous – which means that skaters like Bourque will unfortunately be relegated to the minors.


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|"William Colen (Resolved) (JIRA)" <j...@apache.org>
|[jira] [Resolved] (OPENNLP-450) Add additional context support to POS Tagger
|Sun, 18 Mar 2012 15:00:46 GMT
[ https://issues.apache.org/jira/browse/OPENNLP-450?page=com.atlassian.jira.plugin.system.issuetabpanels:all-tabpanel ] William Colen resolved OPENNLP-450. ----------------------------------- Resolution: Fixed Now the POS Tagger supports additional context. > Add additional context support to POS Tagger > -------------------------------------------- > > Key: OPENNLP-450 > URL: https://issues.apache.org/jira/browse/OPENNLP-450 > Project: OpenNLP > Issue Type: Improvement > Components: POS Tagger > Affects Versions: tools-1.5.3 > Reporter: William Colen > Assignee: William Colen > Fix For: tools-1.5.3 > > > Some applications would benefit from having additional context support in POS Tagger. For example, I could improve the model accuracy by using output from Name Finder. > The change would be to include a field String[][] additionalContext to the POSSample, and modify the code to allow the POS Tagger use it during training and runtime. -- This message is automatically generated by JIRA. If you think it was sent incorrectly, please contact your JIRA administrators: https://issues.apache.org/jira/secure/ContactAdministrators!default.jspa For more information on JIRA, see: http://www.atlassian.com/software/jira
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Smart Tips For Uncovering Professionals
Local Attorneys Can Help You
David Aylor represents clients of different magnitude. Since the year 2006, they have been representing clients in different areas of law. They have attorneys who are specialized in different parts of the law. If you have a matter that falls within their scope; they are more than willing to help you.
They provide criminal defense services. There is high level of anxiety among criminally prosecuted clients as they don not know the outcome of the proceedings. Criminal cases are high profile, and the sentence can be life changing. Their expert attorneys are willing to help you put a strong defense. They accord you high respect and treat all your confidentiality to the letter. They have represented several past clients with positive impacts. Let them have a view of your case regardless of the circumstances that surround the accusations. The lawyers will to their best and see how your case can be handled best.
DUI defense is part of the practice areas of David Aylor law firm. These cases can affect your standing and can even affect your employability. Do not let the stars be added to your driving licenses before the lawyer puts up a defense for you. Once you get them; they will always be a headache. The license looks terrible, and if you have too many, your lines may be cancelled. You will get positive results by hiring a competent lawyer.
Cases of personal injury are on the rise today. A victim of personal injury can bear a lot of challenges. These challenges would not be there if the faulty party behaved responsibly. It is for this reason that her person ought to be made to bear the burden of its action or inaction. The David Aylor attorneys will fight hard for your rightful compensation. It does not matter what was your responsibility, as long as there was a party at fault, you should be compensated.
The workers compensation law was enacted to protect the rights of the employees. The attorneys are fully aware of the changes that employees face and which makes them forego their compensation when injured at the workplace. If you have been injured in your place of work, consult the lawyers for a free case evaluation. They also represent car accident victims. A motor accidents will leave a heavy toll on your health and financial status. With a competent motor accident lawyer, you will get the right compensation for the motor accident.
David Aylor Law attorneys will hold your hand as you fight justice at these trying moments. They will handle all facets of the representation or defense to ensure that you get justice. You can hit the website of David Aylor law firm to learn more about them.


text: Situations Advertising has journeyed through three decades of building brands that resonate with the Indian consumer. Be it the iconic Ujala campaign, Everest Kitchen Spices, or Yardley Fragrances, the company believes in advertising that works in the marketplace and not just in the mindspace. Here's a clean, minimal and crisp website for an agency that means business and is here to stay.


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Today i am banned by whatsapp. i think this is the first of kind of case. They banned me for violating terms of whatsapp.
Most Helpful Girl
you were selling something on there?00|00|0
what was the last thing you did on there?
maybe you called some one bad word or send a nude pic?
nope. no this is not a case with me. i dont do such things on whatsapp. i will tell the reason after i receive all the opinion here. and there is nothing naughty or sexy in this story. and whatsapp not ban some one from doing that. as with the new whatsapp feature chat between to person is not traced by anyone not even by whatsapp. so chill!
Most Helpful Guy
So, it was an error? On your opinion?00|00|0
not an error. but we can see that one can be banned by whatsapp too like facebook or gag.


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Ironville pendant
$442.38 usd $663.5724.80" W × 72.80" H × 24.80" D
Product Info
|Item ID:
|The Ironville ceiling lamp is sophisticated geometric perfection; as slim gold finished metal rods continous wrap to create a an elegant chandelier with four single candelabra lights. Bulbs not included, Bulb sold seperately, Max Watt 25 W, Size E12, Type B35. UL approved and listed.
Shipping & Returns
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text: Pakistan Navy Sailor Jobs Batch C-2017 Female Medical Technician Vacancies: Pak Navy is offering jobs for male and female candidates. Female can apply their job application form for Medical Branch as Female Medical Technician FMT. Male applicants can submit their Pak Navy job online application form for different branches. Pakistan Navy latest jobs 2017 total numbers of vacancies, eligibility criteria, ... Read More »


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The stock climbed 33 percent to 114.75 pence (R19.58) a share, the highest since March 1.
The rand dropped 0.9 percent versus the dollar and has slid more than 9 percent since President Jacob Zuma ordered Finance Minister Pravin Gordhan home from a roadshow before firing him in a cabinet reshuffle.
Lonmin, which raised about $400 million (R5.48 billion) in a life-saving rights offer in 2015, burned through 70 percent of its cash in the final quarter of last year as a low platinum price combined with falling production from its biggest shaft.
The recent rand weakness makes further fundraising less likely, because it lowers the miner's costs, said Arnold van Graan, a Johannesburg-based analyst at Nedbank Capital.
A basket of platinum group metals priced in rand, an indicator of Lonmin's revenue, has climbed 11percent since March 24. The miner pays most of its costs in the local currency.
Read also: Lonmin shares slump after weaker output results
"Lonmin is the platinum stock most geared to the rand, so it's been helped by recent weakness in the currency," Van Graan said.
"As the rand-PGM basket price climbs, the cash burn should decrease, reducing the possible need for external funding."
Short interest in the stock has steadily increased in the past few months, rising to 8.7 percent of shares outstanding as of Tuesday from 2.7 percent on January 31, according to data compiled by IHS Markit.
"The high level of short interest in the stock is creating additional volatilit," Van Graan said.


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"Here are the best nine books I read in March and April with an interesting mix of fiction and non-fiction, adult and children's books." Read more
Search results for tag: Little Women
The Greatest Love Story Ever Told, 30 to 21 – Your votes counted.
February is the month of love at Booktopia, and to celebrate we're counting down the 50 Greatest Love Stories Ever Told, as voted by you. After thousands of votes, the poll closed on Sunday and we'll be counting down the top 50, with the top 10 being announced at Midday on Valentine's Day. So sit back and enjoy the great works that made it to 30-21 in your voting. And don't forget to scroll dow... Read more


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Countryside admits it sold houses with 'doubling' clauses
News comes after Taylor Wimpey's £130m writedown related to controversial leasehold contracts
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Not a Member? Get access to HR news and resources that you can trust.
HR professionals share their advice for minimizing worker stress and boosting retention.
Is your employee handbook ready for the changing world of work? With SHRM's Employee Handbook Builder get peace of mind that your handbook is up-to-date.
Virtual SHRM-CP/SHRM-SCP Certification Prep Seminars kick off September 12 and fill up fast!
Expand your influence and learn how to become an effective leader. Join us in Phoenix, AZ | OCTOBER 2 - 4, 2017
Employer-provided financial and compensation benefits such as flexible spending accounts, tuition assistance and incentive bonus plans are decreasing, according to the
2014 SHRM Employee Benefits Survey from the Society for Human Resource Management (SHRM).
However, the shift to defined contribution retirement savings plans and Roth 401(k) savings plans continues, with only 24 percent of organizations reporting that they offer defined benefit pension plans that are open to all employees.
Among other personal finance-related survey results:
The survey, released annually, polled 510 randomly selected HR professionals. It examines more than 300 benefits.
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Will 3Com Deal Funnel U.S. Secrets to China?
Lawmakers Raise Questions About Bain Capital's Buyout Pact After Approval of Chinese Firm's Stake
Would a private-equity firm founded by Republican presidential contender Mitt Romney fork over sensitive secrets to the Chinese?
It might sound like the plot of a Jason Bourne movie, but it actually is the question shadowing concerns voiced by several U.S. lawmakers over Bain Capital Partners LLC's pending $2.2 billion buyout of 3Com Corp.
Bain's decision to allow China's Huawei Technologies Co. to take a minority stake in 3Com...


text: Performance Commercial flooring spaces require a high performing product. The product must be able to withstand the harshest kind of traffic, whether from people, desks, chairs or other items used in offices, schools or hotels. It must be able to retain its color and pattern against beating sunlight, coffee spills or other stains. In short, it must look as new in a few years as it did the day it was first installed. For more than 40 years, Zeftron has been delivering more to specifiers and end users of commercial carpet products. That's because Zeftron nylon is a premium nylon 6 fiber that not only withstands but also exceeds virtually any performance expectation. Performance Certification Zeftron nylon was the first premium branded yarn system in the industry to establish a performance certification program for commercial carpets. The program provides an unmatched level of assurance that the commercial carpet products made with Zeftron nylon meet or exceed all critical performance characteristics required for a specific end use purpose. Read More The Zeftron nylon Performance Certification program is an independent assessment of a carpet's construction specifications and its expected level of performance when subjected to varying levels and types of traffic. Carpets after evaluation are assigned a traffic class rating of either Class I (moderate traffic), Class II (heavy traffic) or Class III (extra heavy traffic). A carpet must meet minimum performance requirements before it is accepted as a Zeftron premium branded style carpet. Once achieving certification, the carpet is the warranted for life for stain removal, colorfastness, wear and static control. Lifetime Warranties Zeftron stands behind its superior performing product with superior warranty protection. In fact, Zeftron was the first in the industry to offer a lifetime stain removal guarantee and lifetime performance warranties. Read More Today, Zeftron offers lifetime performance warranties for: Carpet Wear -- When installed and maintained as recommended by the Zeftron nylon Service Center, the carpet will retain at least 90 percent of its pile fiber for the duration of its useful life (subject to certain limitations) Static Control – When installed and maintained as recommended by the Zeftron nylon Service Center, the carpet will be shock-proof to human beings for the duration of its useful life Stain Removal – Carpets made exclusively with Zeftron nylon solution-dyed nylon yarn, when installed and maintained as recommended, will resist permanent stains caused by spills of all substances for the duration of its useful life Color Stability – Carpets made exclusively with Zeftron nylon solution-dyed nylon yarn and containing only Zeftron approved yarn or carpet treatments, and when installed indoors and maintained as recommended by Zeftron, the carpet will be colorfast against exposure to light and atmospheric contaminants No other fiber brand in the industry can match the breadth of coverage and level of support provided through these warranties. Please contact us for full warranty details Superior Service and Support Zeftron's commitment to a high performing product does not end with Performance Certification and lifetime warranties. Zeftron delivers more. We provide additional services to further ensure the product specified meets the demands of the space in which it will be expected to perform. Read More Our additional services include: Traffic Classifications – Zeftron can help ensure proper carpet selection based on the traffic classification designated after Performance Certification testing, and also assure the carpet's full protection under the appropriate Zeftron warranty Specification Assistance – The Zeftron nylon team of marketing specialists provide assistance to help clarify the art and science of selecting the appropriate carpet for a particular end-use area Proper Carpet Care and Maintenance Support – Zeftron can provide experienced expertise to extend the carpet's value and beauty years beyond the initial installation Technical Support Should unexpected issues arise, Zeftron provides onsite problem-solving technical expertise and product performance assessment to specifiers and end users for the life of the carpet Environmental Consciousness Through Zeftron nylon's 6ix Again program, we will assist to ensure an environmentally responsible alternative to a landfill disposal at the end of the carpet's useful life Solution Dyed Nylon Specifying a commercial carpet with a premium solution-dyed nylon like Zeftron nylon provides numerous advantages, including vibrant coloring and styling possibilities – as well as superior durability and wear. But be aware: Not all solution-dyed yarns are created equal. Read More A solution dyed nylon such as Zeftron has high quality pigments (color) added during the nylon manufacturing process. Infusing color into the nylon itself produces superior colorfastness characteristics and color retention, especially when compared to a "white" nylon, which has its color added later in the carpet manufacturing process. But remember: not all solution dyed nylon is the same. Simply being solution dyed does not mean the carpet is completely resistant to stains, cleaning chemicals or bleaching. Nor does it mean the carpet will possess superior colorfastness properties or retain its color over the long-term due to other atmospheric contaminants. That's why specifying a commercial carpet made with Zeftron nylon is always the right choice. Delivering More Specifiers and end users requiring a high performing commercial carpet product need to look no further than products made with Zeftron nylon. Millions of yards of installed carpet spanning 40 years might be proof enough. But Zeftron delivers more. Zeftron, a premium solution dyed nylon 6 fiber, is further enhanced with unmatched warranties and value-added services. Combined, these advantages provide unequaled assurance and peace of mind to designers, specifiers and end users. It means that carpet made with Zeftron nylon is guaranteed to meet or exceed its intended end use specification for years to come.