text:
|Publication number
|Publication type
|Application number
|Publication date
|May 28, 1996
|Filing date
|Sep 19, 1994
|Priority date
|Sep 19, 1994
|Fee status
|Also published as
|Publication number
|Jane S. Tjia, Brian D. Kelley, Richard P. Northey, C. Michael Philbrook
|Original Assignee
|Genetics Institute, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (22), Non-Patent Citations (4), Referenced by (106), Classifications (18), Legal Events (6)
|External Links: USPTO, USPTO Assignment, Espacenet
The subject invention relates to the field of osteogenic proteins and pharmaceutical formulations thereof. More particularly, the subject invention involves pharmaceutical formulations designed to sequester osteogenic protein in situ and to provide a bioresorbable scaffolding for a time sufficient to allow the protein to induce cartilage and/or bone formation through differentiation of uncommitted mesenchymal cells into osteoblasts.
Osteogenic proteins are those proteins capable of inducing, or assisting in the induction of, cartilage and/or bone formation. Many such osteogenic proteins have in recent years been isolated and characterized, and some have been produced by recombinant methods. For example, so-called bone morphogenic proteins (BMP) have been isolated from demineralized bone tissue (see e.g. Urist U.S. Pat. No. 4,455,256); a number of such BMP proteins have been produced by recombinant techniques (see e.g. Wang et al. U.S. Pat. Nos. 4,877,864 and Wang et at. 5,013,549); a family of transforming growth factors TGF-α and TGF-β) has been identified as potentially useful in the treatment of bone disease (see e.g. Derynck et at., EP 154,434); a protein designated Vgr-1 has been found to be expressed at high levels in osteogenic cells (see Lyons et al. (1989) Proc. Nat'l. Acad. Sci. USA 86, 4554-4558); and proteins designated OP-1, COP-5 and COP-7 have purportedly shown bone inductive activity (see Oppermann, et at. U.S. Pat. No. 5,001,691).
Various attempts have been made at developing formulations designed to deliver osteogenic proteins to a site where induction of bone formation is desired. For example, certain polymeric matrices such as acrylic ester polymer (Urist, U.S. Pat. No. 4,526,909) and lactic acid polymer (Urist, U.S. Pat. No. 4,563,489) have been utilized, but these formulations do not sequester the osteogenic protein for a time sufficient to optimally induce bone formation, and further have been found to erode too slowly for optimal bone formation.
A biodegradable matrix of porous particles for delivery of an osteogenic protein designated as OP is disclosed in Kuberasampath, U.S. Pat. No. 5,108,753. While U.S. Pat. No. 5,108,753 discloses that a successful carder for OP must bind the protein, act as a slow release delivery system, accommodate each step of the cellular response during bone development, and protect the protein from nonspecific proteolysis, no formulations are suggested which contain components that specifically sequester the OP at the site where bone formation is desired.
Ron et al., U.S. Pat. No. 5,171,579 discloses that the average surface area per porous particle is critical to optimize bone formation.
Brekke et al., U.S. Pat. Nos. 4,186,448 and 5,133,755 describe methods of forming highly porous biodegradable materials composed of polymers of lactic acid COPLA).
Okada et al., U.S. Pat. Nos. 4,652,441, 4,711,782, 4,917,893 and 5,061,492 and Yamamoto et al., 4,954,298 disclose a prolonged-release microcapsule comprising a polypeptide drug and a drug-retaining substance encapsulated in an inner aqueous layer surrounded by a polymer wall substance in an outer oil layer.
Yamazaki et al., Clin, Orthop. and Related Research, 234:240-249 (1988) disclose the use of implants comprising 1 mg of bone morphogenetic protein purified from bone and 5 mg of Plaster of Parris. U.S. Pat. No. 4,645,503 discloses composites of hydroxyapatite and Plaster of Paris as bone implant materials.
Collagen matrices have also been used as delivery vehicles for osteogenic proteins (see e.g. Jeffries, U.S. Pat. No. 4,394,370), but collagen frequently causes undesirable antigenic reactions in patients. Therefore, there remains a need for a pharmaceutical formulation capable of sequestering osteogenic proteins at a site where induction of bone formation is desired for a time sufficient to allow safe, effective induction of such bone formation.
In U.S. Pat. No. 5,171,579, it is disclosed that osteogenic proteins can be sequestered at a site where bone inducing activity is desired using autogenous blood, without using antifibrinolytic agents, provided that a porous particulate polymer matrix is incorporated into the formulation. To reduce the preparation time and improve the above formulation's handling characteristics, Applicants have found that it is desirable to fabricate the porous particulate polymer matrix into the form of a fused sponge, which is preferably wettable and slightly malleable. The fused sponge may be made by wetting the porous particulate polymer matrix with a suitable liquid fusing composition. A preferred liquid fusing composition is comprised of 95% ethanol and 0.05% polysorbate or other non-ionic surfactant. The liquid composition may be added to the particles, preferably in a ratio of about 0.75 to about 0.85 liquid:particles (v/v).
In one embodiment, the subject invention provides methods for preparing fused sponges useful as a carrier for osteogenic proteins. In another embodiment, the invention comprises compositions comprising a pharmaceutically acceptable admixture of an osteogenic protein together with a fused sponge formed of a porous particulate polymer, the particles of which have been fused into a fused sponge form using appropriate processing. The formulations may optionally include other protein sequestering agents, particularly cellulosic materials.
The methods and compositions of the present invention are useful for the preparation of formulations of osteoinductive proteins which can be used, among other uses, to promote the formation of cartilage and/or bone, for repair of tissue damage and fractures.
The inventors have found that, by heating porous particulate polymeric material, preferably copolymers of lactic acid and glycolic acid (PLGA), to a temperature above its glass transition temperature fig), it is possible to fuse them together. However, heating PLGA particles alone results in co-polymer chain mobility and ultimately to particle collapse. This leads to a very dense, non-porous structure, which is undesirable. The inventors have surprisingly found that this undesirable effect can be overcome by adding a liquid containing a lower alkyl alcohol (i.e., C1 -C5), preferably ethanol, and heating the composition in a sealed container. Adding an external plasticizer which is a poor solvent or non-solvent for PLGA in a lower alkyl alcohol, such as ethanol, will lower the Tg of the polymer. If the external plasticizer has a low diffusion coefficient in the polymer, the surfaces of the polymer will fuse together before the particle fully collapses.
Accordingly, the present invention comprises processes of preparing fused sponges of porous particulate polymer, which fused sponges are suitable for use as a carrier for osteogenic protein. Generally, the process comprises:
a) preparing a mixture of particles of a porous particulate polymer, a lower alkyl alcohol and a surfactant;
b) heating the mixture of step (a) until the particles form a fused sponge;
c) optionally continuing to heat the mixture for an additional period of time at a temperature of about 45� to 60� C.;
d) optionally cooling the mixture; and
e) drying the mixture in the form of a fused sponge.
In a preferred embodiment, the particles are made of co-polymers of lactic acid and glycolic acid.
The porous particulate polymer matrix component useful in the practice of the subject invention is a polymeric material that can be formed into the fused sponges of porous particles as described below thereby providing in situ scaffolding for the osteogenic protein, while having biodegradable properties allowing for replacement by new bone growth. Examples are polymers which may be useful in the present invention include polymers of amino acids, orthoesters, anhydrides, propylene-co-fumarates, or a polymer of one or more α-hydroxy carboxylic acid monomers, (e.g. α-hydroxy acetic acid (glycolic acid) and/or α-hydroxy propionic acid (lactic acid)). The latter can be employed in its d- or 1- form, or as a racemic mixture, the racemic mixture being preferred. The preferred polymers for use in preparing the formulations of the present invention are copolymers of lactic acid and glycolic acid (PLGA).
When copolymers of lactic acid and glycolic acid are employed (PLGA), the molar ratio of monomers can range from 1:99 to 99:1 depending upon the desired bioerosion lifetime which in turn depends upon the clinical indication being addressed, as more than 50% of either monomer gives longer bioerosion lifetime (slower biodegradation). The molecular weight of the polymer can range from about 1,000 to 100,000 (relative to polystyrene in CHCl3) with 30,000-50,000 being preferred when a 50:50 copolymer is employed. In general, the higher the molecular weight, the slower the biodegradation.
The polymeric matrix component of the subject invention is used in the form of highly porous to hollow (with surface porosity) particles, hereinafter collectively referred to as "porous particles." These porous particles are generally spherical having diameters of 150 to 850 μm, preferably 150 to 500 μm, most preferably 300 to 500 μm. This particle size creates sufficient spacing between particles to allow mammalian osteoprogenitor cells to infiltrate and be positively influenced by (evidenced by an increase in osteogenic activity/bone growth rate) the osteogenic protein.
U.S. Pat. No. 5,17 1,579 discloses that the average surface area per porous particle is critical to optimize bone formation. Specifically, porous particles useful in bone formation according to the present invention should have an average surface area of from about 0.02 to 4 m2 /g. The disclosure of U.S. Pat. No. 5,171,579 is hereby incorporated herein by reference. WO 93/06872 further discloses that it is possible to produce porous particles having the desired surface area by introducing a "porosigen" (composition capable of imparting porosity by increasing particle surface area) into the solution used to produce the porous particles. The disclosure of WO 93/06872 is hereby incorporated herein by reference. It is also possible to control the bioerosion rate by subjecting the porous particles to sterilizing doses of γ radiation. The higher the γ radiation dose, the faster the bioerosion. Particles useful herewith have a porosity such that the surface area of the particles is increased about 2-250 fold over the surface area of non-porous particles of comparable size.
A preferred method of production of the porous particles useful in the invention is, generally speaking, a solvent evaporation process comprising dissolving the polymer (in e.g. CH2 Cl2), and adding a porosigen such as NaCl, mannitol or sucrose in solid and/or liquid form. When porosigen is added in solid form, the matrix-porosigen solution takes the form of a suspension. Another preferred method of production of the porous particles useful in the invention is a solvent extraction method, wherein the porosigen is added in liquid form with concomitant homogenization. When porosigen is added in liquid form with homogenization, the matrix-porosigen solution takes the form of an emulsion. With either method, the matrix-porosigen emulsion is added to an excess aqueous solution containing surfactant such as poly(vinyl alcohol) with controlled stirring and temperature. The resultant porous particles are hardened by extracting or evaporating residual solvent, and dried. PLGA particles useful in the subject invention made utilizing 50% NaCl as a porosigen have a surface area of between about 0.2 and 0.6 m2 /g; and particles made using sucrose as a porosigen have a surface area of between about 0.04 and 0.09 m2 /g. PLGA particles useful in the present invention made using liquid porosigen with homogenization have a surface area of between about 0.02 and 4 m2 /g.
Other methods of preparing porous particulate polymer for use in the present invention, in which the above steps can be varied and additional steps can be performed are within the skill of the art.
The liquid useful for fusing the porous particulate polymer to form a fused sponge comprises a lower alkyl alcohol, preferably ethanol, and may further comprise a surfactant. The alcohol is preferably in a concentration of from about 50% to about 100% volume/volume, more preferably from about 70% to about 100%. The most preferred concentration of ethanol is about 95%. Substances other than lower alkyl alcohols may also be useful provided that they have a low diffusion coefficient in PLGA, are able to wet the PLGA particles and are able to create vapor pressure to create voids or pores between the particles.
The surfactant useful in the liquid is preferably a non-ionic surfactant, such as a polyoxyester, for example polysorbate 80, polysorbate 20 or Pluronic F-68. The most preferred surfactants include Polysorbate 80, commercially available as Tween 80. The surfactant is preferably present in a range from about 0 to about 4% volume/volume, more preferably from about 0.01 to about 0.05%.
Preferably the ratio of total liquid:solid during heating, that is the ratio of ethanol and surfactant to particles, is from about 0.70 to about 0.90 volume/volume. It is most preferred that the liquid:solid ratio be in the range from about 0.75 to about 0.85.
The container useful for heating the particles and liquid is preferably made of polystyrene. Other materials known in the art, such as polypropylene, may also be useful containers for heating the particles. Preferably, the container is sealed. The container is preferably in the shape and size which is desired for the fused sponge, such as rectangular or cylindrical. Alternatively, the fused sponge may be cut to a preferred size or shape after formation of the fused sponge.
The container is used as a mold for the formation of a fused sponge. The liquid and the particles may be added to the container in any order or simultaneously. After being added to the container, the liquid and particles are mixed, stirred or otherwise manipulated to ensure even distribution of the liquid and the particles.
The liquid and particles mixture is heated at a temperature and for a period of time sufficient to form a fused sponge structure, preferably from about 1 to about 60 minutes until the particles and liquid reach a temperature of about 45� to 60� C. The heating may be accomplished using any suitable heating means. Heating is preferably accomplished using a circulating water bath. Fusion may be accomplished at a constant temperature, or using a temperature gradient. If a constant temperature is used, it is preferably about 60� C. and the mixture is preferably heated for a period of time suitable to allow fusion of the particles and liquid into a fused sponge, generally, about 1 to 60 minutes, most preferably about 10 minutes. If a temperature gradient is used, the heating preferably begins at a temperature of about 26� C. to about 40� C. and the temperature increased at an acceptable rate, preferably about 1.7� C./minute, until it reaches about 45� to 60� C., most preferably about 60� C. Once the mixture reaches the desired temperature, the container may be removed immediately from the bath. Optionally, heating of the mixture at a temperature of about 45� to 60� C. may continue for an additional period of time from about 1 to about 60 minutes.
The fused sponge may be removed from the container immediately upon removal of the container from the water bath. Alternatively, the fused sponge may be allowed to cool or may be refrigerated, preferably in a 4� C. water bath, for a period of time, preferably for a time of about 5 minutes or more, before removing the fused sponge from the container. The cooling step aids in preventing collapse of the sponge which can occur with slow cooling at room temperature. If necessary, forceps or similar means may be used to remove the fused sponge from the container. After removal from the container, the fused sponge is allowed to dry.
The osteogenic proteins useful with the fused sponges made in accordance with the subject invention are well known to those skilled in the art and include those discussed above. The preferred osteogenic proteins for use herein are those of the BMP class identified as BMP-1 through BMP-12 in U.S. Pat. Nos. 4,877,864; 5,013,649; WO 90/11366 published Oct. 4, 1990; WO 91/18098 published Nov. 28, 1991; WO 93/00432, published Jan. 7, 1993; U.S. Ser. Nos. 08/247,908 and 08/247,904, both filed May 20, 1994; and U.S. Ser. No. 08/217,780, filed on Mar. 25, 1994. The disclosure of the above publications are hereby incorporated by reference. The most preferred is BMP-2, the full length cDNA sequence of which is described in detail in the '649 patent. Of course, combinations of two or more of such osteogenic proteins may be used, as may fragments of such proteins that also exhibit osteogenic activity. Such osteogenic proteins are known to be homodimeric species, but also exhibit activity as mixed heterodimers. Heterodimeric forms of osteogenic proteins may also be used in the practice of the subject invention. BMP heterodimers are described in WO93/09229, the disclosure of which is hereby incorporated by reference. Recombinant proteins are preferred over naturally occurring isolated proteins. The amount of osteogenic protein useful herein is that amount effective to stimulate increased osteogenic activity of infiltrating progenitor cells, and will depend upon the size and nature of defect being treated as discussed in more detail below, such amounts being orders of magnitude less than the amount of porous particulate polymer matrix employed, generally in the range of 1-50 μg of protein for each 10 mg of fused sponge employed and more preferably in the range of 0.5-10 μg protein for each milligram of fused sponge employed (assuming approximately 0.2 g/cc density).
The osteogenic proteins can be utilized in the form of a pharmaceutically acceptable solution (including reconstitution from a lyophilized form). It is optimal to solubilize the osteogenic protein at concentrations of at least about 1 mg/ml, preferably about 2 to 8 mg/ml, so that a pharmaceutically effective amount of protein can be delivered without undue volumes of carrier being necessary. For some applications, concentrations above 2 mg/ml may be desirable. Amino acids having a net positive charge (e.g. net 1+species such as arginine, histidine, lysine and the ethyl esters of glycine and beta-alanine), preferably a net 2+charge (e.g. the ethyl ester of histidine, the methyl esters of lysine and arginine, and agmatine), are useful in this regard. Amino acids having a net zero charge are useful in this regard provided that the positive charge of the compound is sufficiently distant (at least 2-3 CH2 units away) from the neutralizing negative charge (e.g. net neutral species such as gamma-amino butyric acid, beta-amino propionic acid, and glycine-glycine dipeptide). Other solubilizing agents useful herein include poly(sorbate), dextran sulfate, guanidine, heparin, sodium chloride, glutamic acid hydrochloride, acetic acid and succinic acid. For use in solubilizing dimeric BMP, such as BMP-2, 3, 4, 5, 6, 7, 8, 9 and 10 and heterodimers of BMPs such as BMP-2/6 and BMP-2/7, preferred solubilizing agents include arginine and histidine (including esters thereof) and glutamic acid hydrochloride. A preferred solubilizing agent is glutamic acid hydrochloride (HCl). The above formulations may be lyophilized and reconstituted with water, providing for advantages in storage, shipping and stability. A preferred formulation of osteogenic protein at 4.0 mg/ml comprises the following components listed in Table 1:
TABLE 1______________________________________osteogenic protein 4.0 mg/ml (11.42% wt)buffer solution:glutamic acid HCl 0.918 mg/ml (2.62% wt)glycine 25 mg/ml (71.39% wt)sucrose 5 mg/ml (14.28% wt)polysorbate 80 0.1 mg/ml (0.29% wt)______________________________________
The porous nature of the particles useful in the present invention creates sufficient surface area for protein adsorption and increases biodegradation, the desirable extent of both being dependent upon the clinical indication being addressed. Surface area can be measured by any conventional technique. For example, BET surface area analysis can be employed using a Micromeritics ASAP 2000 system, which measures surface area based upon adsorption and desorption of Krypton gas at the surface and within the pores of the solid sample. The unit calculates and prints out the surface area: ##EQU1##
By plotting ##EQU2## the slope being ##EQU3## and the intercept being ##EQU4## the surface area ##EQU5## where N=Avogadro's number and V=molar volume. The amount of sponge used to treat a particular defect will, of course, depend upon the size of the defect being treated, and on the effective amount required to adsorb the osteogenic protein.
Additional optional components useful in the practice of the subject application include, e.g. cryogenic protectors such as mannitol, sucrose, lactose, glucose, or glycine (to protect the formulation from degradation during lyophilization), antimicrobial preservatives such as methyl and propyl parabens and benzyl alcohol, antioxidants such as EDTA, citrate, and BHT (butylated hydroxytoluene), and surfactants such as poly(sorbates) and poly(oxyethylenes), etc. Of course, the traditional preparation of formulations in pharmaceutically acceptable form (i.e. pyrogen free, appropriate pH and isotonicity, sterility, etc.) is well within the skill in the art and is applicable to the formulations of the subject invention.
In certain uses, the formulations of the subject invention may be used in combination with various bone cements, including erodible bone cements such as poly(propylene-co-fumarate) and certain hydroxyapatite cements. Also, certain of these uses will utilize bioerodible hardware such as erodible plates, screws, etc. The dosage regimen will be determined by the clinical indication being addressed, as well as by various patient variables (e.g. weight, age, sex) and clinical presentation (e.g. extent of injury, site of injury, etc.). In general, the dosage of osteogenic protein will be in the range of from about 10 to 1000 μg, preferably from about 10 to 100 μg per 100 μL sponge volume.
The osteogenic protein and fused sponges of porous particulate polymers of the formulations may be provided to the clinic as a single formulation, or the formulation may be provided as a multicomponent kit wherein, e.g. the osteogenic protein is provided in one vial and the porous particulate polymeric fused sponge is provided separately.
The formulations of the subject invention provide malleable implants that allow therapeutically effective amounts of osteoinductive protein to be delivered to an injury site where cartilage and/or bone formation is desired. Such an implant may be used as a substitute for autologous bone graft in fresh and non-union fractures, spinal fusions, and bone defect repair in the orthopaedic field; in cranio/maxillofacial reconstructions; for prosthesis integration, especially as a surface coating to improve fixation of prosthetic implants such as hydroxyapatite coated prostheses; in osteomyelitis for bone regeneration; and in the dental field for augmentation of the alveolar ridge and periodontal defects and tooth extraction sockets. When used to treat osteomyelitis or for bone repair with minimal infection, the osteogenic protein may be used in combination with porous microparticles and antibiotics, with the addition of protein sequestering agents such as alginate, cellulosics, especially carboxymethylcellulose, diluted using aqueous glycerol. The antibiotic is selected for its ability to decrease infection while having minimal adverse effects on bone formation. Preferred antibiotics for use in the devices of the present invention include vancomycin and gentamycin. The antibiotic may be in any pharmaceutically acceptable form, such as vancomycin HCl or gentamycin sulfate. The antibiotic is preferably present in a concentration of from about 0.1 mg/mL to about 10.0 mg/mL.]
The following examples are illustrative of the present invention and are not limiting in any manner. Modifications, variations and minor enhancements are contemplated and are within the present invention.
Porous particles of PLGA are produced in accordance with the methods described in International Patent Application WO 93/00050, and U.S. Pat. No. 5,171,579, the disclosure of which is hereby incorporated by reference herein.
A. Sponge Integrity and Wettability
PLGA particles of diameter of approximately 300 to 500 μm are placed in a 1.5 ml Eppendorf vial [internal diameter=8 mm, length=43 mm] with screw caps. 95% ethanol, or 95% ethanol and 0.05% Tween 80 (polysorbate 80) surfactant, is then added and the PLGA particles are packed as tightly as possible by tapping the vial against a hard surface. The vials are capped and immersed in a circulated water bath at 45� C. for 10 minutes. The vials are then removed and decanted. The fused sponge formed is removed from the tube and allowed to air dry overnight. The resulting cylinder is 7 mm in diameter and approximately 40 mm in length. The fused sponge is sectioned into pieces 5 mm in length [approximately 200 μl] and tested for wettability with water and the buffer solution shown in Table 1 above. Rectangular fused sponges are also made using disposable cuvettes as molds. The cuvettes are sealed with two layers of aluminum foil and parafilm. Additional fused sponges are made with 95% ethanol and 0.05% Tween 80 surfactant using 1.5 ml Eppendorf tubes.
Structural Integrity: The fused sponges show a surprisingly high resistance to tensile forces and maintain their shape under high strain. Fused sponges made without any additives have the highest structural integrity. They show a relatively high degree of resistance to tension. Rectangular fused sponges have a higher resistance than cylindrical fused sponges.
Wettability: After being allowed to dry for approximately 24 hours, both the fused sponges with ethanol and those made with ethanol and surfactant wicked quickly and easily. Over time, the fused sponges made with only ethanol gradually lose their quick wicking ability. Those made with surfactant retain their wickability. The cylindrical sponges wick water slightly better than the rectangular sponges. Newly formed cylindrical sponges, both with and without added surfactant, are able to wick approximately 2/3 of their volume per ml of sponge.
B. Process Parameter Optimization:
PLGA particles of diameter of approximately 300 to 500 μm are placed in a suitable container and wetted with 95% ethanol, 0.5% Tween 80 surfactant in a ratio of 0.75 (v/v) ethanol/PLGA particles to form a slurry. The resulting slurry is stirred to ensure even distribution of the particles throughout the ethanol and to remove any large air bubbles which may exist. The mold is sealed and the sponge is heated in a circulating water bath. After heating, the mold is unsealed and allowed to cool before the fused sponge is removed from the mold and allowed to dry. Removal is accomplished by inverting the mold, and depending upon the mold material, either lightly shaking or vigorously tapping the mold against a hard surface until the fused sponge is loosened.
Effect of Temperature:
Fused sponges were made at water bath temperatures ranging from 30 to 75� C. Temperatures are selected to exceed glass transition temperatures of the PLGA particles to ensure that some fusion would take place at the lowest temperature (30� C.). Higher temperatures were examined to see how porosity is affected by more extreme conditions.
Fusion at a constant temperature as well as with a temperature gradient was studied. Two starting temperatures were used for the temperature gradients: 26.4� C. and 40.0� C. Once the molds were placed in the water bath, the temperature was set for 60.0� C. and the water bath was allowed to heat up at an average rate of 1.7� C./min. Once the water bath reached 60�, the fused sponges were either removed immediately or held for 5 minutes before removal. The temperature inside and outside the mold was monitored using a thermocouple in order to better understand sponge and pore formation.
In general, friability decreased as fusion temperature increased. However, deformation of both the PLGA particles and the fused sponge itself was more prevalent as fusion temperature increased. At a temperature of 60� C., sufficient fusion is achieved without completely melting the PLGA particles and with minimum friability. At temperatures greater than 60� C., the PLGA particles lose their structure and overfusion occurs. All of the fused sponges made using a temperature gradient have good porosity. Those removed from the water bath immediately after it reached 60� C. also have excellent sponge shape. The fused sponges held for 5 minutes at 60� C. were noticeably more difficult to remove from the mold. Forceps had to be used to facilitate removal, resulting in deformation of the fused sponge.
Regardless of whether a temperature gradient was used, the inside of the mold approaches the temperature of the surrounding water bath within 2.5 minutes. When 26� C. is the initial temperature, the temperature inside the mold is always slightly below that of the water bath. Acceptable fused sponges are formed using both of these methods.
Effect of Time:
Sponges were heated from 1 to 60 minutes at 60� C. After 1 minute, fusion has already occurred. However, at 1 and 2 minutes, the sponges are prone to a noticeable amount of shedding of particles from the edges when the fused sponge is handled. Friability and shedding noticeably decreases as time is increased. After 1 hour, shedding and friability are minimal. Porosity increases with heating up to 10 minutes. After 10 minutes, further heating only serves to decrease friability.
Removal of fused sponges from the molds becomes increasingly difficult as time increases. At 10 and 15 minutes, a spatula was needed to facilitate removal. However, this does not appear to have a negative effect on the dimensions of the fused sponge when compared to those that did not require the use of a spatula.
Density of the fused sponges was measured at 0,5,10 and 15 minutes. The time of heating has little effect on density. The average density of the fused sponges is 0.17 g/cc.
Effect of Ethanol/Surfactant:PLGA Ratio:
A mixture of ethanol and surfactant, such as Tween 80, is added to the PLGA particles to act as an external plasticizer and to lower the glass transition temperature (Tg). To determine an optimal ratio of ethanol/surfactant:PLGA particles, ethanol/Tween was added to PLGA particles in volume-to-volume ratios ranging from 0.60 to 0.90.
At low ratios (<0.70), sponge deformation tended to occur. This may be due to the possibility that not all of the particles are adequately covered with ethanol. Thus, some particles may have undergone bulk chain mobility and collapse. At ratios of 0.9 or higher, the resulting fused sponges tend to be soft and friable. The greater the amount of ethanol present, the greater the total volume of the slurry and the greater the average distance between particles in the slurry. Thus, at high ethanol volumes, there is less particle-to-particle contact resulting in less, and weaker points of fusion. The liquid to solid (ethanol/surfactant:PLGA) ratio should therefore be in the range of about 0.70 to about 0.90. At a ratio of 0.75, the fused sponge is harder and more rigid than that made at 0.9 and maintains the shape better than at 0.70. Thus, preferred liquid to solid ratios are in the range of about 0.75 to about 0.85.
Effect of Ethanol Concentration:
Sponges were made at 60� C. for 1 hour with ethanol concentrations ranging from 50 to 100% in water. It was found that porosity decreases and sponge deformation increases with decreasing ethanol concentration. However, no appreciable differences in the overall structural integrity existed among the fused sponges.
Effect of Particle Size:
Upon addition of ethanol to the PLGA particles in the mold, a particle density gradient was observed with those particles less than 500 μm. More particles were present at the bottom of the mold than at the top. The gradient was visually less severe as particle size increased. In order to better distribute the particles, molds were tapped for a longer period of time. No density gradient was observed with particles greater than 500 μm in size.
All fused sponges made with particles less than 500 μm were similar in final appearance. Slight porosity gradients were observed with those fused sponges made from 150-250 tμm particles. This is most likely a direct result of the particle density gradient. However, all of these fused sponges (<500 μm) had good porosity, shape and structural integrity.
Fused sponges made with 500-710 μparticles were noticeably less rigid and more friable than those made with smaller particles. These sponges were easily broken when removed from the mold and had slightly greater porosity than the other fused sponges. Because these particles are bigger, there is less contact between particles which could account for the decrease in structural integrity. These fused sponges were also more difficult to remove from the molds intact because of the large pores present. When dry, it was noticed that there was a significant amount of shedding of the particles from the fused sponge when it was handled. This was not the case with fused sponges made from particles less than 500 μm.
Effect of Mold Material:
Fused sponges made with polystyrene as a mold typically contain interparticle pores greater than 300 μm, but contain relatively few pores which are in the range 40-300 μm. Fused sponges made using polypropylene tend to have more interparticle pores in the range 40-300 μm. Because PLGA particles are porous, all fused sponges have intraparticle pores less than 40 μm. Fused sponges made with polypropylene tended to have higher density than those made with polystyrene. For this reason, polystyrene is the preferred mold material, although both polypropylene and teflon may be acceptable.
Removal of the fused sponges from polypropylene and teflon is easier than from polystyrene. Removal from a stainless steel mold was very difficult.
Long-Evans male rats were divided into test groups of five rats each. Each received a subcutaneous implant, 200 μL in size, with a PLGA sponge fused at 60� C. for 60 minutes with rhBMP-2 doses of 0, 10, 20 and 40 μg/100 μL implant. Two alternate formulations of PLGA sponges were prepared which varied the extent of PLGA fusion (underfused=60� C. for 10 minutes; overfused=65� C. for 10 minutes) which were tested at rhBMP-2 doses 10 and 40 μg/100 μL implant to determine of extent of fusion effects the efficacy of the device. After 14 days, the rats were sacrificed and each animal was evaluated for bone formation. As a control, implants comprised of unfused porous PLGA particles in an autologous blood clot matrix, as described in U.S. Pat. No. 5,171,579. The control implants contained 10 μg rhBMP-2/100 μL implant.
The results of the rat ectopic study are summarized in Table 2. The fused sponges of PLGA particles achieved average bone scores between 1.0 and 2.0. No significant differences were found between underfused and overfused devices in either of the doses (10, 40 μg/100 μl rhBMP-2) evaluated. This suggests that the loss of microporosity resulting from overfusion of PLGA does not significantly impact bone formation in this model. All rhBMP-2 containing devices showed evidence of some bone growth on both the periphery and the interior of the implant, although homogeneity of interior bone growth varied among groups.
TABLE 2______________________________________ BEP/Matrix Implant Residual Wet WeightTest Group Cartilage Bone Score (Grams)______________________________________Control/10 0.1 0.85 � 1.6 0.597 � 0.47 0.268PLGA/A/0 0 0 2.4 0.081 � 0.051PLGA/A/10 0.2 1.05 � 2.6 0.122 � 1.07 0.049PLGA/A/20 0.15 1.35 � 2.3 0.259 � 0.85 0.179PLGA/A/40 0.05 1.7 � 2.6 0.174 � 0.82 0.072PLGA/B/10 0.2 1.25 � 2.2 0.129 � 0.92 0.050PLGA/B/40 0.15 2.1 � 2.4 0.269 � 0.88 0.141PLGA/C/10 0.1 1.3 � 2.4 0.220 � 0.92 0.167PLGA/C/40 0 2.0 � 2.6 0.272 � 0.67 0.183______________________________________ Control = Unfused PLGA particles in blood clot matrix PLGA/A = Particles fused at 60� C. for 60 minutes PLGA/B = Particles fused at 60� C. for 10 minutes ("underfused") PLGA/C = Particels fused at 65� C. for 10 minutes ("overfused") Bone and Cartilage scores: Samples were scored on a scale of 0-5 for presence of new bone as per the following table: 5 = 80-100% of the section; 4 = 60-80% of the section; 3 = 40-60% of the section; 2 = 20-40% of the section; 1 = 10-20% of the section; 0.5 = <10% of the section (not significant); 0 = not observed in the section Residual Bioerodible Particles (BEP) or residual matrix within implants were scored on a scale of none (0), low (1), medium (3) or high (5) for the presence of residual BEP in the implant.
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|International Classification
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|European Classification
|Jan 30, 1995
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10 pasos para escribir un guion para animación
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16 Secrets Revealed by Professional ScreenwritersProfessional Screenwriting The authors of previous books on screenwriting had no real industry experience as working screenwriters on produced movies. Or their experience was so old, what they had to teach was not relevant for today's movie industry."16 Secrets Revealed by Professional Screenwriters" is different.The author was able to use his Industry connections to get Successful Professional Screenwriters/and Television
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by S. PriceThe screenplay is currently the focus of extensive critical re-evaluation, however, as yet there has been no comprehensive study of its historical development. International in scope and placing emphasis on the development and variety of screenplay texts themselves, this book will be an important and innovative addition to the current literature.
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by Jack ForbesProfessional Actor/Writer Jack Forbes offers us front-row seats in an enticing array of myriad characters and compelling plot twists. Jack takes us from Big League baseball coach, to homicide detective, murder suspect, rape victims, criminal defense attorneys, private investigator, sketchy eye-witness, clever industrial spy, hapless executive, fighter pilot, female infantry prisoner, mob enforcer
Stuntkohtausten suunnittelu ja kuvaus
by H-P VirkkiKirja esittelee elokuva-alan ammattilaisen silmin toimintakohtausten kuvausten valmistelun, harjoittelun, tekniset apuvälineet ja kuvaukset tavalla, josta saavat vinkkejä niin YouTube-videoiden kuin muidenkin filmien tekijät. Vauhdikkaiden ja todentuntuisten kohtausten toteutukseen ei välttämättä tarvita monipuolista kuvauskalustoa, vaan hyviä ideoita, liikunnallista taitoa ja kuvasilmää.


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Although few days ago Swedish Automobile a.k.a. Saab owner said that it terminated the investment deal with China's Pang Da and Youngman, today we saw a surprising u-turn: Saab was sold to Pang Da and Youngman! That's right, the two Chinese companies bought the Swedish brand for 100 million euros or $142 million.
According to the official statement, the final agreement between the parties is subject to a definitive share purchase agreement between Swan, Pang Da and Youngman.
The whole situation is quite confusing, as last week, Saab said that North Street Capital or the company that recently bought Spyker sports car manufacturer, announced that it will invest $70 million in the troubled company. "There are rumors out this morning that say the deal is signed and so forth, but that is nothing that we can comment on yet,'' said Saab spokeswoman, Gunilla Gustavs.
According to Saab, the MOU is valid until November 15 of this year, but we believe that the Chinese companies will close the deal and Saab will finally be rescued, just like Geely did with Volvo.


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Our innovative, entrepreneurial spirit was born in Linköping in 1983, where ContextVision developed and implemented a super fast image computer, based on long-term research in image processing and the human visual system. Earlier studies of how the human brain processes visual information – with a variety of dedicated cells identifying, defining and classifying image structures – resulted in the development of GOP® methodology, our core technology.
Since then our technology has been developed further to include various technologies for image processing as well as artificial intelligence.
A decision was made in 2015 to adopt a more aggressive R&D strategy in order to assume leadership in medical imaging applications of the latest technologies within machine learning. In line with this, deep learning technology is being developed and combined with our present technologies to create new unique possibilities for image processing and future innovative products.


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14 Home Street Port Macquarie 2444 NSW
This comfortable two bedroom home has a delightful feel and is located in a perfect location for that long holiday feeling. Property features polished floor boards throughout, sunroom, air conditioning, modern kitchen with dishwasher and stainless steel stove, rear entertainment area plus an additional toilet off the laundry. A single lock up garage completes the package. All of this is less than 1 kilometre from the beach and East Port Bowling Club. PETS CONSIDERED. BA
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From the Metroid series' "METROID Other M" comes yet another rerelease of the 1/8th scale figure of the series protagonist, Samus Aran! The figure has her wearing her Zero Suit - the outfit she wears while out on missions, but has the helmet removed to show off her beautiful face!
She looks as cool and calm as ever as she holds her gun in her right hand, ready to take on anything that comes her way. The Baby Metroid that plays an important role in the story is also included along with Samus. It's a figure that really brings out the atmosphere of the Metroid world for fans to enjoy!
- Painted 1/8th scale PVC figure with stand included.
- Approximately 220mm in height.
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A DVD with 30 films by Alexandre Alexeïeff
Four short dance films by Maya Deren
A film by Maya Deren.
Meshes of the Afternoon 1943 16mm 15minAt Land 1944 16mm 16minRitual in Transfigured Time 1945-1946 16mm 16min
'Notes on Marie Menken' documentary + 4 films by Marie Menken
Contenu: 2 DVDs: DVD 1: The 9 restored films of The Magick Lantern Cycle; DVD 2: Extras. + CD + Book Attention! For orders to France, Belgium or Switzerland, please use the link bellow: http://www.potemkine.fr/Potemkine-film/Kenneth-anger-the-magick-lantern-cycle/pa61m3f258.html
A 4-disc Blu-ray/DVD Combo Edition that includes 37 films, restored to pristine high-definition, from some of the foremost experimental filmmakers of the 20th century.
3 films by Maya Deren :Experimental Films, Divine Horsemen, Dance Films


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Clear Blue Fertility Monitor update
View Single Post
March 9th, 2009, 10:03 PM
Mega Super Mommy
Join Date: Dec 2008
Location: Middle TN
Posts: 2,818
Well after getting the "go ahead" from my new doctor, I decided to start using the fertility monitor this month just to kind of have a heads up and head start on figuring out what's going on with my ovaries and if I ovulate.
So far nothing too interesting. I was surprised that it had me start POAS on CD6. I guess I was surprised just because I knew with the PCOS I probably will have a longer than 28 day cycle so CD6 seemed early. But, this is the first cycle with the monitor and it takes a little while to "learn" your cycles.
So from CD6 which was the 5th (actually since the monitor is a day behind my actual counting, I guess it was actually CD7... but whatever), it has said I have "high" fertility, but not "peak." The directions do say that the first cycle is a "getting to know you" and then after that it'll be more individualized and sensitive.
but so far so good. It's a little annoying being told to POAS every morning because even if I get up needing to pee, and turn on the monitor to be told to POAS, it takes a good 5 (maybe more) minutes to get me the results so I'm sitting there waiting when I just wanna go back to sleep! haha
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If the proteins have long been known nutritionists, research allows gradually to better identify their roles in the body and refine the recommended intake. Zoom on the latest advances.
Not to be confused with the meat or fish that contain the proteins are among the energy nutrients as well as carbohydrates (sugars) and fats.
What is a protein?
Every protein consists of the assembly of smaller molecules, amino acids. With digestion, protein ingested with food are broken down into amino acids. Once assimilated, they allow the body to renew body protein. Twenty amino acids can thus be combined, of which 9 are essential. The body can not produce them, they are required to be supplied by the diet.
What are the proteins in the body?
Protein is abundant in the human body: they represent 10 to 12 kilos of the body of an adult male, 250 to 300 g are renewed each day. About 40% of body proteins are localized in the muscles, but they enter the structure of all cells. The bones are thus constituted by a matrix protein (mainly collagen) on which is fixed calcium. Some body proteins have important functions: hemoglobin that can transport oxygen to various organs, insulin and glucagon are the hormones regulating blood glucose (blood sugar), the immunoglobulins or antibodies involved in the defense against infectious agents, digestive enzymes that convert food into digestible nutrients .
Do the children need a lot of protein?
Constituting each tissue, proteins are important nutrients for growing organisms. However, the recommended intakes are not much higher in children and adolescents compared to adults: 0.78 to 0.90 grams per kilogram of body weight per day, instead of 0.83 g (for example, a child weighing 10 kilos needs 8 9 g of protein per day). There are a few years, studies have left thinking that excess protein in childhood obesity favored a few years later. Although this link is now being questioned by researchers, it is not worth giving too much protein for young children, who spontaneously consume more than necessary.
In practice, in infants, breast milk or milk "first age" provides exactly the right amount of protein. Later, the contributions are met by 3 to 4 dairy recommended per day and a small portion of meat or fish of 10 g per year of age: 30 g to 3 years, 60 g to 6 years … From Similarly, pregnant women do not need more than 1 g of protein per kilogram of weight.
He must seniors insist on their protein intake?
From age 60, the recommended intake increases of 0.83 g of protein per kilogram of body weight per day to 1 g and 1.2 g even in people over 75 years. The goal is to slow the physiological muscle loss (sarcopenia), which slowly begins to quarantine but is increasing significantly after 65 years. Between 50 and 80 years, the loss of muscle is about 30% among people who do not practice regular physical activity … with consequences in everyday life: loss of strength and mobility little by little reduced, fall risk in the elderly. Recent studies show that it is possible to slow down this process and even maximize the muscle mass by practicing 1-2 times per week of strength training (strength against a load or force) and having sufficient consumption proteins. Among seniors, take most of the day protein in the same meal would facilitate the protein synthesis of organisme.
Sports they consume too much protein?
To retain more muscle mass than sedentary individuals, athletes (those who practice at least 1 to 2 hours per day, 4-5 times a week) need a little more protein: 1.2 g 1.5 g per kilo of body weight per day. Some, especially those who train in strength, tend to consume more, easily double. According to recent studies, these high intakes are not required, training outweighing protein intake to build muscle mass.
We know however that consumption of protein (about 20 g) associated with sugars (40 to 60 g) in the half-hour after workouts promotes muscle mass gain: for example, one large bowl of chocolate milk or 40 cl drinking yoghurt sucré4. The health consequences of excess protein are still imperfectly known, but it is shown to be harmful in people with kidney failure (sometimes unknowingly) 1.
They are the proteins to lose weight?
high protein sachets or Dukan diet, proteins are often being honored in dieting. It is shown to have a satiating effect, that is to say, they minimize hunger pangs from one meal to another and thus they help moderate energy intake. But if high protein diets do lose weight quickly, it is mainly because they provide very few calories. Nutritionists insist that a good intake of protein prevents muscle wasting. What is true, but on condition of not drastically reduce energy intake and practice enough physical activity (at least 30 minutes per day) .
What foods are proteins?
conventionally distinguished animal proteins, present in meat, fish, eggs, dairy and vegetable proteins in significant proportion in oil seeds (almonds, sesame …), legumes (soybeans and pulses ) and cereals.
The former are a bit better assimilated and / or better with essential amino acids. Eat balanced by pins of the National Health and Nutrition Programme (3 dairy and 1-2 servings of meat or fish per day, bread or starchy food at each meal) largely can meet the recommended intake of protein.
Should we rely on plant proteins?
The "semi-vegetarianism," which involves eating vegetarian part-time, is a growing trend. Eat meat less often is beneficial to the planet, livestock is a major consumer of water and great purveyor of greenhouse gases. From a nutritional point of view, make a greater emphasis on plant proteins allows to consume less saturated fat (including excess promotes increased blood levels of bad cholesterol, or LDL) that are associated with animal protein in meats, cold cuts and cheeses. We can, for example, once a week, replace meat or fish by pulses, whose protein quality will be optimized by the association in cereals (each type of protein compensating for the weaknesses of the other point view of the essential amino acids): rice and lentils, chick peas and semolina (couscous), pasta and white beans (minestrone). Tofu "soy cheese" or plants steaks can also replace the time meat to time, provided they do not consume more than two soy products (juice, "yoghurt", tofu, tempeh …) day , because of their intake of phytoestrogens, compounds similar to féminins6 estrogen.
Finally, for those who might be tempted to replace dairy products with vegetable juice, be aware that only the soya juice has a similar protein content of cow's milk (rice juice, hemp, almond … are very poor ) and that these products do not provide calcium unless fortified.


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As the ending credits of 2015 draw near, it's time for Take 2 movie critics Rebecca Kivak and Tamara Dunn to list their best and worst of the year in film. The duo sound off on best film, worst film, screen surprise, favorite character and hidden gem.
Best film
Tamara's Take
Here is my take on the year in movies.
Best film
"Spotlight": This should be the template on which tense dramas should be based. Many in the audience know about the Catholic priests scandal before watching "Spotlight," but few know about how hard the Boston Globe crew worked to get the story. With a top-notch cast of Michael Keaton, Mark Ruffalo and Rachel McAdams, the actors create suspense where the viewer didn't know it could exist.
Honorable mention: "Creed," a great spinoff to the classic "Rocky" series; "Brooklyn," a realistic romantic film.


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500 Macy Gift Card [US]
$500 Macy's Gift Card [ US ]
What about $500 to spend on Macy's? I think it sounds amazing when holidays are coming. If you don't try you never win!
E-Mail Based Sweepstakes. Confirm and Continue. DO NOT USE PROXY/TOR Browser.
Tags: macy gift card, us sweepstake, usa gift cards, win macy gift card


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GraphicRiver Cake 4427814
Category: Stock Vector | Comment: ( 0 )
GraphicRiver Cake 4427814
Stock Vector \ Objects \ Food
Isolated \ Image \ Illustration \ Holiday \ Meal \ Painting \ Vector \ Sweet \ Glaze
Stock Vector - Cake 4427814 by ded_Mazay @ GraphicRiver
Stock Vector \ Objects \ Food
Graphics Files Included: JPG Image, Vector EPS; Layered: No; Minimum Adobe CS Version: CS.
Cake. Isolated object. No transparency or gradients used. JPG and EPS vector files.
Avatars Aliens; Money Laundering; Teacher; Striking Ammunition; Spray Can ; Slug; Skeletons in the Closet.
Keywords: birthday, cake, candle, cartoon, cream, dessert, drawing, eating, feast, food, fun, glaze, holiday, illustration, image, isolated, meal, painting, pie, sweet, vector.
Download Link


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Kendall Jenner had her first kiss in the shower.
The 20-year-old model had instigated a game of Truth or Dare so she could get close to her "biggest crush", but struggled to find privacy to lock lips with her classmate because her younger sister, Kylie Jenner, kept "spying" on them.
She revealed on her app: "The kiss was with a kid I had THE biggest crush on -- shout out to Chad Wilcox! I crushed on him for so long.
"We were at my mom's house and played Truth or Dare -- of course, the point of that was to kiss each other. ;)
"Kylie was so nosy, so we kept going to secret places like the trampoline --and Kylie kept spying and, like, popping out of trees, lol.
"We were like, 'Go away!' and she was like, 'What are you doing?!?' She followed us to the living room, then the bedroom.
"We even locked her out with a chair under the doorknob and she found her way in. So annoying. She kept saying, 'You're being shady!' "
The pair eventually hid themselves away in the bathroom - because it had a lock.
She continued: "We ended up in my bathroom because it was the only room with a lock! Finally, we got to the Truth or Dare kissing.
"[My friend] Jessica and I didn't want to do it in front of each other so Chad and I went in the shower behind the curtain. And that's how the magic of my first kiss went down! ;).(sic)"


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Alice and the Fly Paperback
by James Rice
This is a book about phobias and obsessions, isolation and dark corners. It's about families, friendships, and carefully preserved secrets. But above everything else it's about love. Finding love - in any of its forms - and nurturing it.
Miss Hayes has a new theory. She thinks my condition's caused by some traumatic incident from my past I keep deep-rooted in my mind.
As soon as I come clean I'll flood out all these tears and it'll all be ok and I won't be scared of Them anymore.
The truth is I can't think of any single traumatic childhood incident to tell her.
I mean, there are plenty of bad memories - Herb's death, or the time I bit the hole in my tongue, or Finners Island, out on the boat with Sarah - but none of these are what caused the phobia.
I've always had it. It's Them. I'm just scared of Them. It's that simple.
- Format: Paperback
- Pages: 336 pages
- Publisher: Hodder & Stoughton General Division
- Publication Date: 27/08/2015
- Category: Modern & contemporary fiction (post c 1945)
Other Formats
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Showing 1 - 1 of 1 reviews.
Review by 4everfanatical
This book is very mid-range for me.

It is a story told largely through the eyes of Greg, a troubled teen who battles with mental illness. Greg's wealthy family are largely emotionally absent, and Greg is the complete opposite of popular in his school. Told through Greg's journal entries, the reader gets an intimate portrayal of his sad and depressing life. The only ray of positivity is his crush on a girl from school -Alice. In between Greg's journal entries, the reader is treated to a police interview transcript about Greg referring to a tragic incident.

The actual event was pretty anti-climatic for me and so this book didn't really have the 'wow' factor. It is pretty easy read, and the story is interesting but it wasn't a book that amazed me.
Also by James Rice | View all


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This is a split board - You can return to the Split List for other boards.
You're browsing the GameFAQs Message Boards as a guest. Sign Up for free (or Log In if you already have an account) to be able to post messages, change how messages are displayed, and view media in posts.
There is really no way to screw up unless you are extremely clumsy.
True, with the exception of the motherboard CPU socket pins, which are extremely fragile and need to be treated with great care. Most things only fit one way though, so unless you force something into a place it's not meant for, it's not going to break.
My movie and gaming room: http://imgur.com/a/IjWr1
My pets: http://imgur.com/a/P9VGU
reincarnator07 4 years ago#22
We've spent a lot of time and given no recommendations and asked no questions...
TC, what is your budget? Do you need mouse/keyboard/monitor/speakers/any other peripherals?
Fan of metal? Don't mind covers? Check out my youtube and give me some feedback
sauruschamp1 4 years ago#23
A good build that will play most games on high (but might struggle maxing a few) is:
Kingston ValueRAM 8GB (2x 4GB) Desktop Memory Kit DDR3
Gigabyte GeForce GTX 760 OC 2GB Video Card
Intel Core i5 3570K Unlocked Quad Core LGA 1155 3.4GHz (3.8GHz Turbo) Processor
OCZ ZT Series OCZ-ZT650W 650W Modular Power Supply
Gigabyte GA-Z77X-D3H Motherboard
Lite-On iHAS324 Internal Optical Drive - 24x12x8x /24x12x6x/12x + 48x24x48x, DVDRW, CDRW - SATA - Retail
Samsung 830 Series 128GB 2.5" Solid State Drive (SSD) - SATA III - 520MB/s Read & 320MB/s Write - 2xnm Toggle DDR NAND - 3-Core ARMv9 MCX Controller (MZ-7PC128B)
Noctua All-In-One Performance Multi-Socket CPU Cooler
Just add a HDD of the size you want (ie 1TB or 500Gb or whatever floats your boat) and a case (this is really personal choice) and your OS (basically a windows 7 cd).
If you have extra money change from a gtx760 to a gtx770 just depends how much you want to spend. Also use pcpartpicker.com it will tell you if any parts are incompatible (usually the only thing you have to worry about is that the motherboard has the right socket for your cpu)
Xbox GT: Zarosian SteamID: Zarosio 3DS ID: 1848-2118-1570
Always forgive your enemies; nothing annoys them so much. - Oscar Wilde
Fade2black001 4 years ago#24
daemon_dan posted...
Nicodimus posted...daemon_dan posted...Well... he's kinda dumb. The parts should be under warranty, and it's not hard to say "The pins were bent when I got it" and RMA it.
Biostar doesnt do this. I know this first hand.
We're Americans! We don't quit just because we're wrong.
We just keep doing the wrong thing until it turns out right.
ZeraphLordS 4 years ago#25
ecollegepc will charge a small fee to put together the components of your choosing, while still retaining the savings and efficiency of building your own.
take a good build from this topic, and get them to build it for you; they're this board's recommended staple.
post a budget and I'm sure someone can help you out :)
The best course of action is to just get the information you need, then get out while you're still alive. - destroy everything on GameFAQs
Marioface5 4 years ago#26
THEskeksis posted...
we are not that kind of computer savvy. are there people who will custom build one for you? i know, how noob, but seriously we arent the building computer types.
It's understandable to be intimidated by the thought of building your first computer, but it really is a lot easier than it seems. As long as you're careful, you should be fine. There really aren't that many steps to it, and as someone else said there are tons of useful guides and videos to help you do it. Maybe you have a friend or family member that can help?
If you want to get into PC gaming, it's important to understand how your own computer works. Not only will building one let you get better parts of significantly less money, but it will also make you far more familiar with how your computer works and what exactly is inside of it. Should you ever need to fiddle with your hardware, you'll definitely want to have been the person who put it together in the first place.
It's your choice, of course, and if you're really that uncomfortable with the idea then I guess that's understandable. But I really can't emphasize enough how much better off you'll be if you take the time to learn how to build a computer and put yours together yourself. If you're undecided, I suggest looking up some of those guides and videos and see what you think of it all. I hope whatever you choose to do works out for you.
The world gets weirder every day.
PSN: Marioface5
I would suggest something like mine(see my sig) but switch the cpu with an Fx 8350 and the SSD with a Samsung 840, or switch motherboard and go intel, Asrock Z77 Extreme4 with an i5 3570k, you can also go with an HD7970 instead of 7950
Asrock 970 DE3/U3S3 \ M12 II Bz 520w / Ballistik 12Gb DDR3 / Ocz 64Gb SSD / HDD 500Gb / XFX HD 7950 3Gb BE Oc / FX 8150 / Hyper Evo 212 / Haf 932 Advanced
If you want a custom built PC but don't want to build it yourself ecollegepc.com is a great site. I have several friends who have ordered from them and all have had great experiences.
FX-6300 @ 4.3ghz | GTX 760 | 970 extreme3 | 8GB DDR3 1600
SteamID: gsf4lyfe
THEskeksis (Topic Creator)4 years ago#29
thaks for all of the helpful posts, im not new to here so i knew at least one person would be "that guy". but we will watch the videos =]
the types of games again are:
Final Fantasy XIV: a realm reborn
Elder Scrolls Online
possibly Battlefield 4
and pretty much whatever looks good that will be coming out. like i said before we are looking for quality over price. we are also going to start looking for places that will custom build for us. that way we can ease into getting into PC gaming and start tinkering around gradually.
really appreciate the help guys/girls!
any products that we should stay away from? top shelf products that are always out doing themselves?
I used to be addicted to soap, but I'm clean now...
THEskeksis (Topic Creator)4 years ago#30
Marioface5 posted...
THEskeksis posted...we are not that kind of computer savvy. are there people who will custom build one for you? i know, how noob, but seriously we arent the building computer types.
how inspiring!! we are so used to console gaming (plug and play) that we didnt think twice about how much we should look into haha now to do some homework....
I used to be addicted to soap, but I'm clean now...
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Young writers collect 2003 Robin McNair Prizes
LONDON - first posted 25 November 2003 - 670 words
Teenage pupils were at the Westminster parliament building today to collect cash prizes which they won by writing essays about the status of early embryos.
The Robin McNair Prize commemorates one of Britain's leading fighter pilots in the second world war. After the war Squadron Leader McNair fought for the rights of the unborn through his membership of the Society for the Protection of Unborn Children (SPUC). The competition is jointly sponsored by the SPUC Educational Research Trust and the McNair family.
The first prize was presented by Mr Alistair Burt MP, parliamentary private secretary to Mr Michael Howard, the Conservative leader, and a member of the Evangelical Alliance's council. Mr Burt said: "As a war-hero, Squadron Leader McNair was daily encountering matters of life and death which is perhaps why, after the war, he was so devoted to the pro-life cause. Our modern society desperately needs to re-acquaint itself with these vital issues, though I expect that change won't come from my generation but rather from the generation to which today's prize-winners belong. I congratulate them, as well as the McNair family for inspiring this award."
John Smeaton, SPUC's national director, paid tribute to Mr Burt's work in the 1980s when he worked to oppose the legalisation of embryo research.
Mr Gerald Soane, grand president of the Catenian Association, presented the second prize. He said: "Our organisation is very pleased to be associated once more with the Robin McNair Prize. Many of our members have teenage children and we welcome SPUC's educational work. The young need to be informed about moral issues, and not just from an amoral, worldly standpoint."
Mr Duncan McNair, son of the late squadron leader, said: "Our family is pleased to associate my father's name with this initiative to educate school pupils in crucial moral matters. I have been hugely impressed with the quality of the entries, particularly the winners' essays which matched the high standard shown by previous years' winners. We wish these young people every success in their future studies and hope that the Robin McNair Prize will be another positive entry on their university applications and CVs."
Judges for this year's competition included Mr Gary Streeter, Conservative MP for Devon south west, and Countess Josephine Quintavalle, founder of Comment on Reproductive Ethics. Countess Quintavalle said: "I was very impressed to see how these young people grappled with such important moral issues and wrote about them in such a clear and interesting way." Mr Streeter presented the prizes last year.
The first prize of £250 was won by Miss Helen Price, aged 19, of Railway Terrace, Penrhiwceiber, Mountain Ash, Rhondda Cynon Taf, who is studying mathematics at Swansea university. She used to attend Mountain Ash Comprehensive School and plans to put the money towards the cost of a computer for her studies.
Second prize of £150 was awarded to Miss Rachel Howard, nearly 17, of Ryehill Avenue, Chesterfield, Derbyshire, who is in the lower sixth form at Repton School, Derby, and used to attend St Elphin's School, Darley Dale, Matlock, Derbyshire. She had to cut her first draft of 2,500 words so that it did not exceed the competition's limit. Miss Howard did most of the writing in a single day but then edited the work over the succeeding weeks. She is taking four A levels and may read geography at university.
Third prize of £100 was won by Miss Rachel Irvine, 16, of Oakgrove Integrated College, Gransha Park, Londonderry, County Derry.
The winners' schools will also receive £250, £150 and £100 respectively.
Miss Price and Miss Howard attended the ceremony, as did Mr David Church, head of religious studies at Miss Price's school. The ceremony was in the Attlee Suite of Portcullis House which is one of the parliamentary buildings at Westminster.
Details of the competition, which is now in its fourth year, can be found at www.spuc.org.uk/mcnair. Essays are of between 1,000 and 1,500 words and the topics change from year to year.
Source: SPUC


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What are some behaviors that one expresses when guilty or approached about something they are embarrassed about. I think for me I'll try to avoid the person....lol
Exaggeration, close-gared gestures (keep arms close to the body), avoiding eye contact. I actually wrote a hub about this: hubpages.com/hub/Detecting-Lies-Neuro-Linguistic-Programming
Yup. I agree, the manarisms, the lil eccentricities in someones behaviors. But there are alot of other things also, exaggeration, deflection, denial, minipulation of the conversations to change the subjects, and the infamous freudian slip among many others. Probrably more than we could list... But all are relative to that individual also. I have seen people that just look like they're lying, and others you would trust with your life and might not live to regret it.So not all, are aplicable to all.
In my personal experiences, I've seen people try to hide their guilt preemptively. When someone is being overbearing towards you trying to figure out what you're hiding when there's nothing going on, they might be trying to hide something themselves.
It's called projection in psychology.
You know that old saying, "The woman doth protest too much"? It applies to men also. I've also heard an involuntary physical reaction is looking up and to the right...
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- Description: This super sexy amazingly hot hunk gives a stripper style dance show while wear a very sexy skin tight black skimpy dress. This video is a solo homemade video. The best homemade striptease solo amateur video on the Net!!
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