text:
|Publication number
|Publication type
|Application number
|Publication date
|Jun 23, 1992
|Filing date
|Jan 31, 1990
|Priority date
|Jan 31, 1990
|Fee status
|Publication number
|Lewis L. Lanier, Joseph H. Phillips
|Original Assignee
|Becton, Dickinson And Company
|Export Citation
|BiBTeX, EndNote, RefMan
|Non-Patent Citations (4), Referenced by (16), Classifications (10), Legal Events (6)
|External Links: USPTO, USPTO Assignment, Espacenet
This invention relates to novel complex co associated with the CD3ζ protein on CD16- NK cells, and more particularly relates to a purified form of the complex, a gene encoding the complex and monoclonal antibodies to the complex. Methods of using the monoclonal antibodies to detect cells bearing this novel complex also are disclosed.
NK cells (or "Natural Killer" cells) are lymphoctyes found in the peripheral blood, in bone marrow and in other lymphoid tissues. NK cells are not considered to be T cells because they do not rearrange or productively transcribe the α, B, γ or δ genes of the T cell antigen receptor complex (or "TcR"). NK cells have ability to kill and lyse certain tumor and virally infected cells. Unlike T cells, however, the killing activity of these cells is not restricted by the major histocompatiblity complex (or "MHC"). NK cells also secrete interferons and cytokines that regulate immune responses and hematopoiesis.
Morphologically, NK cells usually are large granular lymphocytes with prominent azurophilic granules. NK cells also may be identified by the expression of certain markers or structures on their cell surface. These structures can be detected by monoclonal antibodies which have been prepared by conventional methods against them.
The prototypic structures (or "antigens") expressed by the majority of NK cells are CD16 and CD56. (The CD or "Cluster Designation" is an international standard used in the field to define a particular antigen by its molecular weight, cell type distribution and other criteria. The most recent Workshop to cluster antigens and thus antibodies thereto was held in Vienna in February 1989. The most recent list of cluster designations has been published in Journal of Immunology, 143:758 (1989)).
CD16 is an antigen of approximately 50-70 kD and is an Fc receptor for IgG. Besides NK cells, CD16 is expressed on granulocytes, activated macrophages and a minor subset of T cells. Functionally, the binding of immune complexes (e.g. antibody coated target cells) or anti-CD16 monoclonal antibodies to CD16 on NK cells causes a rapid increase in intracellular Ca++ and inositol trisphosphate (or "IP3 ") generation with subsequent transcription of lymphokines and triggering of cell mediated toxicity.
Two highly homologous genes encode CD16. CD16-I encodes a phosphatidylinositol(PI) glycan linked glycoprotein expressed by granulocytes, whereas CD16-II encodes a transmembrane-anchored glycoprotein expressed on NK cells and on activated macrophages. The expression of CD16 II on the surface of COS-7 cells (a monkey kidney tumor cell line, ATCC CRL 1651) requires co-transfection with the γ subunit of the high affinity IgE receptor or the CD3ζ chain.
Monoclonal antibodies can be raised against CD16 by conventional techniques such as those described by Kohler and Milstein. See Nature, 256:495 (1975). For example, mice can be immunized with large granular lymphocytes (or NK cells) isolated from human peripheral blood. Spleen cells from mice so immunized then can be hybridized to a mouse plasmacytoma cell line, such as P3-X63 (ATCC TIB 9), in the presence of a agent such as polyethylene glycol. Clones then can be screened for reactivity with NK cells. Anti CD16 monoclonal antibodies are commercially available from Becton Dickinson Immunocytometry Systems (or "BDIS") as Anti-Leu 11.
CD56 is an antigen of approximately 145-220 kD. It is expressed on essentially all resting and activated NK cells, on approximately 5% of T cells but not on granulocytes, monocytes or B cells. Anti-CD56 monoclonal antibodies can be prepared as above using the KG-1a cell line (a human hematopoietic cell line, ATCC CCL 246.1) as an immunogen. Commercially, an anti-CD56 monoclonal antibody is available from BDIS as Anti-Leu 19.
Not all NK cells, however, express CD16. Similarly, there are NK cells that express quantitatively less CD56. Using anti-CD16 and anti-CD56 monoclonal antibodies which have been fluorescently labelled by conventional techniques with chromophores such as phycoerythrin and fluorescein isothiocyanate in combination with a flow cytometer, three distinct subsets of NK cells are found: 1) a small population of CD56bright, CD16negative cells; 2) a small population of CD56bright, CD16dim cells; and 3) a major population of CD56dim, CD16bright cells ("Bright" and "dim" refer to the fluorescence intensity of the cell stained with a labelled monoclonal antibody. The cells that express the most antigen will be "bright", those that express less antigen will be "dim" and those that express no antigen will be "negative.") These populations are referred to as CD16-, CD16hu + and CD16++ NK cells, respectively. The CD16++ population comprises ˜15% of total peripheral blood lymphoctyes (or "PBL") while the CD16+ and CD16- populations comprise less than 1% of PBL. It is thought that CD16+ cells eventually become CD16++.
Besides expressing CD16 and CD56, NK cells also express a variety of other antigens including CD2, CD7, CD11b. CD38, CD45R, CD18 and the p75 subunit of the interleukin 2 receptor (or "IL 2R"). A minor population of NK cells also express low surface density CD8. None of the CD16-, CD16+ or CD16++ NK cell populations express on their cell surface CD3 γ, δ or ε or react with antibodies that preferentially bind the α/B or γ/δ subunits of TcR. Additionally, none of the populations react with antibodies against T cell associated antigens CD4, CD5 or CD28, the B cell associated antigen CD19 or the monocyte associated antigen CD14.
There are, however, differences between the three populations of NK cells. For example, 50-60% of the CD16++ NK cells express CD57, but neither the CD16+ nor the CD16- NK cell populations generally express CD57. On the other hand, both the CD16- and CD16+ populations express a high surface density of Leu-8 (a 70-80 kD antigen that the BDIS monoclonal antibody Anti Leu-8 reacts with on the majority of PBL, thymocytes and B cells and which was prepared using peripheral blood T cells as an immunogen) but only 20% of CD16++ NK cells also express Leu-8. Further, CD16++ NK cells express no CD25 but the CD16+ and CD16- NK cell populations express detectable levels of CD25.
Functionally, CD16- NK cells isolated directly from blood have little cytolytic activity against human tumor cell targets such as K-562 (ATCC CCL 243). CD16+ and CD16++ NK cells both show cytolytic activity against NK sensitive tumor cell targets with the CD16++ NK cells showing a greater activity. On the other hand, CD16- and CD16+ NK cells have a greater proliferative response to IL-2 than do CD16++ cells.
Taken together, the several NK cell populations appear to comprise a continuum of differentiation in the peripheral blood with the CD16- population being the most immature and the CD16++ population being the most mature. Accordingly, in adoptive immunotherapy, such as that described in U.S. Pat. No. 4,607,007, it may be more appropriate to use CD16- or CD16+ NK cells to achieve a greater effect.
Apart from this characterization of NK cells, other work has been done to determine how NK cells recognize a target cell in the absence of the MHC. As previously described, T cells can function in a cytolytic manner but usually do so only in the context of the MHC. It is believed that the recognition function by the T cell is mediated through the TcR but that signal transduction to turn on the cells cytolytic function occurs through the co associated CD3 structure via the PI pathway.
The T cell antigen receptor is composed of an α/B or γ/δ TcR heterodimer that is non-covalently associated with CD3. CD3 is a complex of at least five subunits, designated CD3γ, δ, ε, ζ and η. CD3γ, δ, ε, ζ are monomeric proteins encoded by genes closely-linked on chromosome 11, whereas CD3ζ is on chromosome 1 and expressed as a disulfide linked homodimer or a heterodimer in association with CD3η. Antigen induced T cell activation results in tyrosine phosphorylation of CD3ζ. It is believed that efficient signal transduction via the CD3/TcR complex requires expression of CD3ζ.
Because NK cells lack surface CD3 γ, δ and ε and TcR, there has been interest in determining how recognition occurs and how signal transduction occurs. Recently, it has been reported by Anderson et al., Nature, 341:159 (1989), that CD3ζ is expressed on NK cells. Anderson et al. further reported that there were apparently novel structures associated with CD3ζ. In FIG. 4, they show 12 kD, 60-70 kD and 80-90 kD proteins all associated with CD3ζ. Surprisingly, however, it has been discovered that the 60-70 kD structure is not novel but is, in fact, CD16. Thus, there is little information to suggest through what mechanism signal transduction occurs in NK cells that lack CD16.
The present invention comprises a novel structure co-associated with CD3ζ on NK cells. The structure comprises a protein complex of approximately 50 kD and which may be isolated in purified form.
The present invention further comprises monoclonal antibodies directed against the protein complex and methods of using the protein complex as an immunogen to produce monoclonal antibodies against it.
The present invention still further comprises a method of detection of hematopoietic cells bearing the protein complex.
The present invention also comprises a method for killing tumor targets by NK cells using an effective amount of a monoclonal antibody or antibodies against the protein complex in vivo.
FIG. 1 comprises a series of autoradiographs of acrylamide gels of granulocytes (A) and NK cells (B C) that have been labelled with 125 I and subsequently immunoprecipitated with either a rabbit Ig control, anti-CD3ζ antibody or anti-CD16 antibody.
FIG. 2 comprises an autoradiograph of an acrylamide gel of the 50-70 kD bands taken from FIG. 1B which then were reacted with various amounts of S. aureus protease V8.
FIG. 3 comprises an autoradiograph of a two dimensional acrylamide gel (1st dimension non reducing, 2nd dimension reducing) of NK cells labelled with 35 S methionine and cysteine and subsequently immunoprecipitated with a rabbit Ig control (A) or anti-CD3ζ antibody (B).
FIG. 4 is an autoradiograph of an acrylamide gel of CD16+ NK cells (A) and CD16- NK cells (B) labelled with 125I and immunoprecipitated with a rabbit Ig control, anti-CD3ζ and anti-CD16.
FIG. 5 is an autoradiograph of an acrylamide gel of CD16- NK cells labelled with 125 I and immunoprecipitated with a rabbit Ig control and anti-CD3ζ under (A) non reducing and (B) reducing conditions.
The present invention relates to a novel protein complex designated p50. To elucidate this structure, the relationship between CD3ζ and CD16+ on NK cells was examined first.
The IL-2 dependent NK cell line was used as a source of NK cells having the phenotype CD3ε-, CD56+. (This cell line was established in cell culture conditions as described by Yssel et al., J. Immunol. Meth., 72:219 (1984)). As a control, granulocytes obtained from peripheral blood also were used. The cells were labelled with 125 I as described in Lanier et al., J. Exp. Med., 165:1076 (1987). (Note that CD3ζ does not contain tyrosine residues exposed on the cell surface; therefore, CD3ζ should not be labelled efficiently by the 125 I ) After labelling, the cells were lysed in 10 mM Tris and 150 mM NaCl (pH 7.2 containing 1% digitonin (CalBiochem), 1 mM PMSF (phenyl methyl sulfonyl fluoride) and 20 KU/ml aprotinin (Sigma)). These conditions were used to preserve the non covalent associations of membrane protein complexes.
Nuclei were removed by centrifugation at 13,000�g for 5 minutes. Cell lysates were precleared three times with 10 mg Pansorbin™ (Protein A, CalBiochem-Behring) coated with saturating amounts of rabbit anti-mouse Ig serum. Antigens were immunoprecipitated with Pansorbin™ precoated with control rabbit Ig, rabbit anti-CD3ζ (provided by L. Samelson, National Institutes of Health) or anti-CD16 (clone CLB-FcGranl, provided by P. Tetteroo and T. Huizinga, Amsterdam).
The structure precipitated by the anti-CD3ζ antibody further was eluted from the immunoabsorbant by incubation for 3 hours at 4� C. in 200 μl of 50 mM Tris and 150 mM NaCl (pH 8 containing 1.5% NP 40 (Nonidet P 40) and protease inhibitors). The eluted material was re immunoprecipitated with Pansorbin™ coated with control Ig or anti-CD16. See FIG. 1C.
Samples of all cell lysates were denatured in sample buffer containing 5% 2 mercaptoethanol, and then were analyzed by sodium dodecylsulfate polyacrylamide gel electrophoresis (or "SDS-PAGE", 10% acrylamide). An autoradiograph of the gels then was made.
Referring to FIG. 1A, samples of granulocyte lysates immunoprecipitated with rabbit Ig (or "C"), anti-CD3ζ or anti-CD16 were run. As expected, CD16 was bound by the anti-CD16 antibody in the lysates while neither the control nor anti-CD3ζ lanes showed any reactivity.
Referring to FIG. 1B, samples of the NK cell lysates immunoprecipitated with rabbit Ig, anti-CD3ζ or anti-CD16 were run. The lanes of both the anti-CD3ζ and anti-CD16 bound lysates show a structure between 50-70 kD. Referring to FIG. 1C, it is evident that the 50-70 kD structure that co-precipitated with CD3ζ is CD16 and not some other novel structure as reported by Anderson et al., supra.
To confirm this point, the 50-70 kD bands from FIG. 1B were cut from the lanes. They then were separately digested for 1 hour with 0, 5 or 25 μg of S. aureus V8 protease (Sigma) and analyzed by SDS PAGE (15% acrylamide). Referring to FIG. 2, the bands cut from the anti-CD3ζ and anti-CD16 lanes cut from FIG. 1B both show essentially identical peptides further indicating that the 50-70 kD structure non covalently bound to CD3ζ is CD16.
To confirm that CD3ζ was, in fact, being expressed by the NK cell line used, the NK cell line described above was metabolically labelled for 18 hours in methionine and cysteine free RPMI-1640™ (a cell culture media (Gibco)) containing 0.2 mCi/ml 35 S-methionine and cysteine (Trans 35 S label, ICN), 25 mM HEPES (pH 7.4), gentamicin and 5% dialyzed fetal calf serum (or "FCS"). Cells were lysed in 50 mM Tris and 150 mM NaCl (pH 8 containing 1% NP-40, 1 mM PMSF and 20 KU/ml aprotinin) for 20 minutes at 4� C. Nuclei were removed as above. Lysates were immunoprecipitated with either control rabbit Ig or anti-CD3ζ. Samples were analyzed by two dimensional diagonal SDS PAGE (1st dimension non-reducing; 2nd dimension reducing; 12.5% acrylamide).
Referring to FIG. 3A, it can be seen that the rabbit Ig does not precipitate any off diagonal structure from the lysates. In FIG. 3B, however, the characteristic CD3ζ disulfide linked homodimer was elucidated at 16 kD.
Given the role of CD3ζ in activation through the TcR, it is likely that CD3ζ is involved in signal transduction via transmembrane anchored CD16. The requirement for the CD3ζ homodimer for expression of CD16 in NK cells is analogous to the requirement for the γ homodimer for the expression of the high affinity IgE receptor (FcR) present on mast cells and basophils It is noted that the transmembrane segment of γ FcR subunit is highly homologous to the CD3ζ transmembrane segment suggesting that the interaction site is between residues in the transmembrane segments of CD3ζ and CD16. The results indicate that CD3ζ or a family of homologous structures may co associate with membrane receptors involved in signal transduction in several different cell types. The issue that is left open, however, is how signal transduction is achieved in CD16- NK cells.
CD16- NK cells were isolated from normal peripheral blood by purifying CD16-, CD56+ lymphocytes. The cells then were labelled with 125 I as above and lysates were prepared as above. The lysates then were immunoprecipitated with rabbit Ig, anti-CD3ζ and anti-CD16 as above. The lysates then were subjected to SDS-PAGE (12.5% acrylamide) as above.
Referring to FIG. 4B, it can be seen that neither the control nor the anti-CD16 lanes show any specific reactivity. The anti-CD3ζ lane, however, shows the 16 kD structure of CD3ζ but also shows a 50 kD complex not previously seen. This complex has been named p50. In FIG. 4A, the p50 protein does not appear indicating the probable absence of the complex on CD16+ NK cells. FIG. 5 confirms that the p50 protein complex is stable under both reducing (B) and non reducing conditions (A).
To purify the p50 protein complex, labelled CD16- NK cells, such as those described above, can be used. Using an anti-CD3ζ antibody, the p50 protein can be isolated by co-precipitation. The protein then can be eluted from anti-CD3ζ, concentrated and purified. Alternatively, lysates of CD16- cells can be prepared and the p50 protein can be eluted from gels of anti-CD3ζ co-precipitates. The p50 protein complex can be separated by using conventional protein sequencing methods, as described in Protein Sequencing: A Practical Approach, IRL Press, (Findlay et al., ed.s, 1989), for example, and oligonucleotide proteins constructed from the derived polypeptide sequence in order to isolate the gene for p50.
In another embodiment, mRNA corresponding to the p50 protein can be isolated from CD16- by density dependent centrifugation. Once the mRNA is isolated, cDNA can be prepared from it by using reverse transcriptase under the appropriate conditions and inserted into an appropriate phage or plasmid vector. The cDNA encoding proteins of the p50 complex can be isolated by several strategies including, but not restricted to, screening bacterial expression systems (e.g., λgtll, Stratagene) or mammalian expression systems (e.g., CDM8, Invitrogen) using antibodies against the p50 protein complex. See generally Molecular Cloning: A Laboratory Manual (2nd ed.), Cold Spring Harbor Laboratory Press (Sambrook, Fritsch and Maniatis eds., 1989).
Once the sequence of the cDNA is known, the amino acid composition of the p50 protein complex can be determined.
There are several options available for making an antibody that will bind to the p50 protein. In one embodiment, p50 complex proteins purified by any of the methods set forth above can be used to immunize mice, rabbits, rats or other animals to raise antisera to p50 complex. In the more preferred embodiment, monoclonal antibodies that will bind to the p50 protein can be prepared. In this embodiment, mice are immunized with cells bearing p50 or more preferredly with purified p50. The spleens are removed after a period of time and spleen cells are fused with an immortal mouse plasmacytoma cell line, such as SP2/0 (ATCC CRL 1581) or P3X63 in the presence of a fusion agent, such as polyethylene glycol. The resulting hybridomas then can be screened for reactivity of their supernatants with purified p50 or cells expressing the p50 complex. Hybridomas producing monoclonal antibodies that bind to p50 can be cloned for further analysis of their reactivities.
The antibodies produced as described above that bind to p50 can then be used in a diagnostic assay to identify and isolate p50 bearing cells. In one method, the antibodies can be labelled with an enzyme, such as horseradish peroxidase, a fluorescent dye, such as a phycobiliprotein, or a radioisotope, such as 125 I . Cells binding to the labelled antibody can then be detected by conventional methods, such as flow cytometry or gamma counting. In a more preferred method, the antibody is a monoclonal antibody and the label is a fluorescent dye. The fluorescent dye can be directly conjugated to the antibody by means such as those described in U.S. Pat. No. 4,520,110 for phycobiliproteins, U.S. Pat. No. 4,876,190 for peridinin chlorophyll complex, or if the monoclonal antibody is made in a mouse it can be detected using a labelled goat or rabbit anti-mouse antibody as a second step reagent. In the preferred method, cells that express p50 are detected by means of a flow cytometer, such as that described in U.S. Pat. No. 4,599,307 or 4,745,285.
Given that CD16+ NK cells can be activated using antibodies against the CD3ζ co associated CD16 protein, it also may be possible to use an antibody that binds to p50 to stimulate such activity in CD16- NK cells either in vitro or in vivo. The dose of administration of the antibodies can be in an amount sufficient to activate the p50 bearing cells. In this method, it is preferred that the antibodies be monoclonal and more preferably be chimeric for in vivo applications, such as those described in U.S. Pat. No. 4,816,397 or GB Pat. Appl. No. 2 188 638 A (published Oct. 7, 1987). It is to be appreciated that the antibody also can be used to isolate p50 bearing cells from peripheral blood. In this method, IL-2 (or another immune stimulator, e.g., interferon) can be used to activate the isolated cells in vitro before being returned to the body. Alternatively, peripheral blood containing p50 bearing cells can be activated in vitro using the antibodies.
All publications and patent applications mentioned in this specification are indicative of the level of ordinary skill in the art to which this invention pertains. All publications and patent applications are herein incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference.
It will be apparent to one of ordinary skill in the art that many changes and modifications can be made in the invention without departing from the spirit or scope of the appended claims.
|Allison et al., Ann. Rev. Immunol. 5:503 540, 1987.
|Allison et al., Ann. Rev. Immunol. 5:503-540, 1987.
|Clevers et al., Eur. J. Immunol. 18:705 710, 1988.
|Clevers et al., Eur. J. Immunol. 18:705-710, 1988.
|Citing Patent
|Filing date
|Publication date
|Apr 6, 1992
|Mar 22, 1994
|The United States Of America As Represented By The Department Of Health And Human Services
|Evaluation and treatment of patients with progessive immunosuppression
|Mar 17, 1993
|Sep 17, 1996
|United States Of America As Represented By The Secretary Of The Department Of Health And Human Services
|Evaluation and treatment of patients with progressive immunosuppression
|Sep 16, 1994
|Nov 18, 1997
|The Wistar Institute Of Anatomy And Biology
|Human cytotoxic lymphocyte signal transduction surface protein (P38) and monoclonal antibodies thereto
|Oct 28, 1994
|Mar 24, 1998
|Eastman Kodak Company
|Polypeptide surface marker for cells
|Sep 16, 1996
|Mar 30, 1999
|The United States Of America As Represented By The Department Of Health And Human Services
|Evaluation and treatment of patients with progressive immunosuppression
|Aug 6, 1997
|Aug 31, 1999
|Sigma-Aldrich Co.
|Method of identifying cells with polypeptide surface marker
|Oct 24, 1996
|Oct 12, 1999
|Schering Corporation
|Purified mammalian NK antigens and related reagents
|Oct 8, 1999
|Apr 30, 2002
|Sigma-Aldrich Co.
|Purification of recombinant proteins fused to multiple epitopes
|Jan 13, 1995
|Jul 4, 2006
|The United States Of America As Represented By The Department Of Health And Human Services
|Janus family kinases and identification of immune modulators
|Oct 3, 2001
|Aug 22, 2006
|Sigma-Aldrich Co.
|Purification of recombinant proteins fused to multiple epitopes
|Aug 1, 2005
|Feb 10, 2009
|United States Of America As Represented By The Secretary Of The Department Of Health And Human Services
|Janus family kinases and identification of immune modulators
|Jan 15, 2009
|Mar 8, 2011
|The United States Of America As Represented By The Department Of Health And Human Services
|Janus family kinases and identification of immune modulators
|Dec 9, 2005
|Apr 27, 2006
|Sigma-Aldrich Co.
|Purification of recombinant proteins fused to multiple epitopes
|Aug 1, 2005
|Feb 1, 2007
|Chua Janet B Y
|Phosphor based on a combination of quantum dot and conventional phosphors
|Jan 15, 2009
|May 28, 2009
|Government Of The United States Of America, As Represented By The Secretary, Department Of Hea
|Janus family kinases and identification of immune modulators
|Jul 21, 2011
|The United States Of America, As Represented By The Secretary, Dept., Of Health & Human Services
|Janus family kinases and identification of immune modulators
|U.S. Classification
|435/7.21, 435/7.24, 436/547, 436/548
|International Classification
|Cooperative Classification
|European Classification
|Apr 16, 1990
Owner name: BECTON, DICKINSON AND COMPANY, A CORP. OF NJ, NEW
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Effective date: 19900406
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|Jan 18, 2000
|Maintenance fee reminder mailed
|Mar 22, 2000
|Surcharge for late payment
|Mar 22, 2000
|Fee payment
Year of fee payment: 8
|Dec 19, 2003
|Fee payment
Year of fee payment: 12


text:
Staying at Ca' Maffio is not just a holiday. The characteristic building with a few rooms for guests takes on another dimension when rooms are located in a unique and special environment, where hospitality and warmth of a home are combined with an excellent background.
As the B&B Ca' Maffio, located in the Natural Park of Sile, Veneto, provides a holiday in contact with nature, including walks along the lush sides of the River Sile. With its three bedrooms and one suite, Ca' Maffio is a B&B of charm, it is intimate and private, and was originally a monastery of the fourteenth century. The building looks like a stately home with a beautiful porch and the interior furnished in period style.
From the B&B Ca' Maffioyou will have easy access to Venice and Treviso, and you will be able to enjoy these cities which are rich in history, architecture and arts. Far from the beaten track, you can reach Venice by river, admiring along the way, the old mills and patronal villas.
The owners Nicolò and Giuliaare glad to meet you at Ca' Maffio with a warm welcome and a well prepared room waiting for you. They will also happilyprovide you with information on the surrounding places to visit, the traditions, and of all the activities you can experience in their beautiful land.
Altrocanto is another very interesting Boutique Hotel, which is located in Farindola, in the Gran Sasso National Park in Abruzzo. In this wonderful natural setting, a stay atAltrocantosimply means to enjoy an unrefurbished view of nature and history; of forests, waterfalls and caves, and discover ancient sites such as the villages and shrines.
Thomas and Marco will welcome you in their beautiful B&B with five rooms furnished with wooden furniture, made with warm colours. If you love reading this is the perfect place where you can relax with your favourite book in the different corners of the house, or choose between the volumes kept in the rich library available while sipping tea and coffee offered to guests. Wi-Fi is always available for those who like to stay connected to the internet for entertainment or for any job, even during the holiday.
Altrocanto was restored from an old furnace from the 900 which was then transformed into a house in the '20s, and this charming B&B it still has the look of a real home, both in decor and in its organization. A house surrounded by nature is sensitive to the environment as well asenergy saving and the owners have specifically chosen a biomass boiler and photovoltaic panels to produce electricity, thus combining the love of tradition with modern technological tools. Collecting rainwater to water the garden is another important feature of this B & B perfectly placed in the surrounding nature.
If you ever decide to explore nature at the National Park of Gran Sasso, you are encouraged to face it with a spirit of adventure and also explore the surrounding areas. You will find interesting hiking trails, horseback riding or biking, easily accessible from Altrocanto.
Choose a B&B of charm for your vacation.

subset: commoncrawl


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Both of my rear electric windows in my 02 Explorer are giving out. My brother n law has a 01 Explorer that he bought wrecked for the engine. Will the window motor/calbes from the 01 work for my 02? I understand they may not be meant to swap out but if it is possible it will save me from having to buy new window motors & cables for mine!
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Thursday, May 16th, 2013 AT 9:43 AM


text:
Palm tree on George Smathers Beach in Key West, Florida, USA
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The Honorable Rob Bishop
Subcommittee Chairman
The Honorable Joseph J. Heck (R-NV)
Member of Congress
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Prime Insurance Company
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Adventure Partners, LLC
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Nantahala Outdoor Center
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American Outdoors Association
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Idaho Outfitters and Guides Association
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Western Rivers Guides
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Aaron Bannon
National Outdoor Leadership School
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John Duncan
Telluride Outside
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The focus of this hearing is to examine obstacles put in place by the federal government that put more costs and burdens on small businesses and limit public access to public lands. Subcommittee Members will hear testimony from stakeholders who have firsthand experience in how the federal government's restrictive policies are not only standing in the way of recreation, but hurting job creation and economic growth. In many Western states, the federal government owns a majority of the land, and the ability to protect public access is crucial to their economic livelihoods.
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Today, the House Committee on Natural Resources held an oversight hearing on improving management and utilization of federal lands in and around North Las Vegas. Chairman Rob Bishop (R-UT), Rep. Crese......
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Yesterday, the Government Accountability Office (GAO) released a report on the Bureau of Land Management's (BLM) accounting and management of natural gas emissions on onshore federal lands. Committee ......
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3D Bioprinting Market Outlook 2022: Acute Market Reports
3D Bioprinting Market Overview:
The worldwide 3D Bioprinting market volume was evaluated at 481 million US dollars in year 2014. It is predicted to observe a substantial expansion and growth. Also it will receive maximum attention from healthcare researchers during the estimated period due to growing occurrence of chronic ailments resulting in tissue and organ transplantations combined with growing life span of persons and restricted amount of donors of organs.
View Full Report with TOC @ http://www.acutemarketreports.com/report/3d-bioprinting-market
Modernization in techniques and inclusion of information technology within the healthcare sector, growing old age population base and enhanced research & development efforts made for the development and growth of this sector are predicted to serve the long term interests of this sector. All these factors are going to have high impact on the growth of the 3D Bioprinting market encouraging the expansion of the market. As on year 2015, many educational institutes, universities as well as research institutes have begun examining the future prospects of bio printed implants, tissue reconstructions and organs. For example, the Herriot Watt University of Scotland made a 3D bio printer capable of printing the pluripotent stem cell cultures applied for testing the medicines and drugs.
Yet another instance displaying the considerable growth of the market is the great current achievement and breakthrough in the field of thyroid gland transplantation in mice by using the 3D Bioprinting technique for treating ailment of hypothyroidism. If the outcomes are positive, the firm will begin testing the technique for using it for human beings in next few years.
3D Bioprinting Market By Technology:
Technologies used in 3D Bioprinting sector includes syringe based 3D Bioprinting, magnetic levitation, inkjet based 3D Bioprinting and laser based 3D Bioprinting. Magnetic levitation along with syringe based 3D bioprinters held the largest market share in year 2014. The laser technique is predicted to observe the rapid growth in future. Also, syringe based technique is predicted to get higher adaptation in future.
Browse All Reports of This Category @ http://www.acutemarketreports.com/category/ict-market
The next generation magnetic levitation 3D bio printers are predicted to resolve more than eighty percent of the recent failure problems along with improving of precision and speed. Magnetic levitation printers find its applications in human cell generation, toxicity screening, drug response management and human cell generation.
3D Bioprinting Market By Application:
Main applications examined in the study includes biosensors, dental, Bioinks, medical, food and animal product Bioprinting as well as consumer & personal item testing. The medical section is forecasted to lead the global 3D Bioprinting market with more than thirty percent of market share in year 2022. It is predicted to cover the regeneration of implants, organs, tissues and prosthetics along with producing of medical invoice.
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A medical pill is the recent breakthrough in the global 3D Bioprinting market. It is predicted to observe high growth during the forecasted period. This expansion and growth can be ascribed to the growing requirement of drugs and the benefit of fast as well as cost effective manufacturing of bio drugs by using 3D Bioprinting. As per the different market experts, the pharmaceutical sector is predicted to grow manifolds thereby raising the size of medical invoice production.
3D Bioprinting Market By Region:
Sub Continent of North America is predicted to lead the global 3D Bioprinting market in terms of total income during the estimated period. It held more than 36% of the total market share in year 2014 and is predicted to grow at a CAGR growth rate of 10.3%. Presence of a large old age population as well as chronically sick population within the region combined with good healthcare infrastructure, efficient health care services, rise in per capita income and high patient disposable levels of income are the main drivers encouraging the growth of the 3D Bioprinting market in this region.
Further, intensive research & development events recently carried out within this region of North America are predicted to be important growth aspects during the estimated period.
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Space-based solar energy jumps the shark — or fries the shark Star-Wars style
This is a three-fold update on my earlier piece "PG&E signs first-of-a-kind space solar power deal with Solaren. Why?" I will reprint an email sent to the media from physicist Marty Hoffert that begins:
The PG&E deal is a scam. Pure and simple. We don't need to study it in detail any more than one needed to study Bernie Madoff's investment scams.
And I will reprint the remarkably lame response Solaren gave me to the key question of lifecycle greenhouse gas emissions.
But first, let's start with this amazing story in Wired, "Hurricane-Killing, Space-Based Power Plant" based on Solaren's 2006 patent for "altering weather using space-born energy" (see inset figure from patent below, click to enlarge).
[Note to Wired: You are now running difficult-to-close pop-up ads that are likely to cost you readers unwilling to spend more than 2 seconds trying to see your actual content.]
Many readers of the original post were concerned the device could be used as a weapon. Not so far-fetched an idea now — at least no more far-fetched than Solaren's plan to weaken or alter hurricanes from space.
So let's call this Solaren's first self-inflicted wound on its own credibility.
Second would be the email response I got from the company in response to my question "Does somebody have a lifecycle CO2 or GHG emissions calculation per kWh given the fuel needed to launch this stuff?" Cal Boerman, Director Energy Services for Solaren, replied:
Solaren plans to use launch vehicles (Atlas V/Delta IV Heavy Class) that primarily use liquid hydrogen and liquid oxygen for fuels. The resulting emissions are water. These fuels are formed via electrolysis. The Wikipedia definition is: Electrolysis of water is the decomposition of water (H2O) into oxygen (O2) and hydrogen gas (H2) due to an electric current being passed through the water. Solaren assumes the electricity used for this process was generated from clean resources.
Therefore the lifecycle environmental impact per kW-hr is negligible. Also, we do not use solid rocket motors so there is no added pollution from them.
Hope this Helps
Well, It helps me understand how little Solaren has thought about this important issue. Electrolysis is good for generating pure hydrogen, but it is incredibly electricity intensive (duh) as is making liquid hydrogen for transport. Presumably a lot of this is done at night when electricity is cheap — if someone can find information on who exactly makes hydrogen for NASA, I'd love to see it. All I could find is this 2002 article that says it is done near New Orleans using " technology that releases large amounts of carbon dioxide into the atmosphere." Plus they lose a lot of hydrogen through evaporation from the trucking. And of course the trucking uses a lot of fossil fuels.
Making hydrogen from renewable-based electrolysis would probably triple the cost of the fuel. And if Solaren really thinks it can cut launch costs by the factor of 10 or more needed to make this entire effort viable, then it can't be tripling the cost of the fuel.
Finally, here is the full email from Hoffert:
The PG&E deal is a scam. Pure and simple. We don't need to study it in detail any more than one needed to study Bernie Madoff's investment scams. There's no way to do this any more than there is a way to get 12% return on investment consistently regardless of the economy. Didn't stop investment in Madoff and it may not stop investment in this harebrained scheme.
There's no way to get 200 Megawatts from orbit with microwave beaming by 2016 from private sector investment. The infrastructure to do it efficiently with microwaves requires huge structures in orbit and in-space assembly by robots. This is very far from existing technology. Microwaves are the wrong way to start a space solar power business. What we can do in a few hundred kilowatts with laser beaming to PV modules on Earth in a five year time frame because there's no in-space assembly needed and single-launch vehicles could likely do it. This could realistically lead to a buildup of a viable orbital and power industry. Even so, we will need major up-front money to test the idea from the feds. The promoters of the PG&E deal idea say they'll provide a thousand times more power and do it all from the private sector. Might as well say we're ready to go to the Moon or Mars with private sector financing. The physics of this is very well understood by the research-active SBSP community.
Too bad, because when it all unravels it will be a major setback for space solar power. Ken [Caldeira], this is very much like your experience with the company that wants to get rid of CO2 in seawater by a proprietary process that violates basic chemistry. Their CEO says he has special insider knowledge to do this, and so does the company pushing this space solar power deal. His defense it that he took many companies public. These ideas get as far as they do most because people making business decisions about alternate energy are often scientific illiterates. There are real technological and scientific hurdles, showstoppers, that is; and there are often potential effective technical and scientific approaches around them.
The problem is not knowing the difference. It's a much a disaster to overestimate the prospects for near-term profit based on flawed physics as to underestimate the longer-term potential of a new technology based on the opportunities that physics does provide. As Richard Feynman sagaciously observed, "You can't fool Mother Nature." If only we didn't have to deal with those idiotic Homo sapiens primates inhabiting this planet. All very depressing because I'm a strong advocate space solar power technology.
Marty Hoffert
Professor Emeritus of Physics
New York University
'Nuff said.
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12 Responses to Space-based solar energy jumps the shark — or fries the shark Star-Wars style
I'm not so sure weather alteration is far-fetched. The upper atmosphere is very thin, and there's not much mass up there. Large fluxes of microwaves or radio waves beamed into the upper atmosphere can have large heating effects on the atmosphere. So while we may not be able to stop hurricanes, we may be able to steer them. The huge phased array antenna HAARP in Alaska has been rumored to be doing such experiments for years, I think.
Far-fetched or not, it does not appear to be a good idea. Aren't we seeing enough unintended consequences, these days? Do we need to add more?
The way to do space based solar, in the long run, I think, is with a ballistic launcher or space gun from the earth or especially a mass driver from the moon. Electricity cost to orbital velocity from a ballistic launcher is only a dollar or so a pound, and lunar materials could be routinely launched using a mass driver for less than that. Once the ingots of alumnium, steel, silicon, or containers of water or whatever are up in orbit, then they could be processed into useful forms, I think, by automated facilities launched more gently by rocket. The biggest shortage of materials for orbital space colonies would be volatiles, I think, which could be supplied from the earth, while lunar materials could supply all the rest.
Here's a reference giving historical background on the idea, which they call a space gun:
Needless to say, development is required. Wikipedia says that orbits would have to be corrected for the projectiles, perhaps using solid rocket thrusters contained in the projectiles, which would increase cost and complexity. Getting the projectile through the lower atmosphere is the biggest challenge.
We can do space colonies and space based solar, in the long run, I think, if we live, and survive what appears to be runaway global warming.
In the short run, this PG&E deal is likely to be some sort of scam, a military scheme to get power to forward deployed troops as Aviation Week reports, a corporate propaganda effort to confuse the issue of global warming, or a true covert weather alteration scheme.
None of these appear to be good ideas or have anything to do with climate change.
We will see an increase in these magic solutions.
This strategic ignorance and deliberate denial of the problem is key to continued heavy carbon consumption. The delusion reads "We can keep up our carbon energy because some sort of technical solution will fix everything."
This is similar to jumping out of the airplane and expecting to make a parachute on the way down. No one has ever done it, but it is sketched out and technically possible.
We already have something that changes the weather, or more accurately, climate if you will. There are 250 million of them in the United States. Even if you clean it up using the latest technology it still spews 19 lbs of CO2 for every gallon of this fuel it consumes. By now you're way ahead of me. It's the car. It's human nature, I suppose, to ignore the things in front of you and the behaviors that contribute to and take from your daily lives and reach for some outlandish idea of why the climate is changing.
As Professor Hoffert points out, this project will need major up-front money, but not all of it will come from the feds. California utilities are allowed to charge rate payers for new technology before it's been implemented, so they have little incentive to invest in technology they know will work. Seconding Richard, PG&E should be spending more time on proven technologies closer to home that we can really count on, not expensive magic fixes in space.
Hydrogen is a highly-refined form of natural gas.
Anyway, the Atlas V burns kerosene for its main stage, plus a little hydrogen in its upper stage. A quick look suggests it burns ~900 t of propellant, of which ~1/4 is carbon. So, five launches release ~4 kt of CO2. If the station delivers 200 MW for twenty years, that's 1.7 TW-hr, so ~2 g_CO2 / MW-hr.
I agree Richard, we'll see an increase in these magical solutions.
It's crazy, we humans buck at the thought of paying 0.1 cent in the dollar for boring measures we know will work, yet get excited at dramatic schemes that are almost certain to cost vastly more.
Are we somehow evolved to prefer big and bold engineering to dull, behind the scenes changes, irrespective the likelihood of success?
"California utilities are allowed to charge rate payers for new technology before it's been implemented, so they have little incentive to invest in technology they know will work."
If true, this ticks me off. As a PG&E rate payer, am I now going to be charged for space solar, rather than earth solar, wind, or small scale hydropower?
Has actual money changed hands, yet? Signing the deal would seem to imply that.
Has this turned into a SciFi blog?
Or has the world gone mad…
Re [Note to Wired: You are now running difficult-to-close pop-up ads that are likely to cost you readers unwilling to spend more than 2 seconds trying to see your actual content.]
If you choose to run the FireFox browser with the Noscript addon you wont have to put up with that junk, and you will be safer surfing too.
(Basically you choose what runs at what site – so there is a bit of customising in th einitial stages – very easy to use though)
Keep up the good work
NASA makes its hydrogen from natural gas which is the cheapest method, if you don't include the cost of releasing large amounts of CO2 into the atmosphere. Using electrolysis to make the hydrogen would be more expensive, but the cost of fuel is an insignificant fraction of the total cost of putting payloads in orbit. At best, it would cost about $15,000 per kilogram to put solar power satellites into geostationary orbit using Atlas V and Delta IV rockets. In practice it costs about two thirds more than that. But the launch cost could be roughly halved by using cheaper Russian and Chinese rockets.
Even with a launch cost of only $5,000 a kilogram, 200 watts of generating capacity per kilogram and a discount rate of 10% the launch cost alone would add about $2.20 per kilowatt-hour to space based electricity. The total cost would be much more than this, making it vastly more expensive than ground based solar. In many countries it would be cheaper to pay people to pedal bicycle generators than to use space based solar power.
"if someone can find information on who exactly makes hydrogen for NASA, I'd love to see it."
I don't know off the top of my head, but you mention that the rockets they're talking about using are the Altas V and Delta IV. Those aren't NASA's rockets; they're United Launch Alliance (consortium of Boeing and Lockheed Martin), with help from the Air Force. That might help track it down.
"Making hydrogen from renewable-based electrolysis would probably triple the cost of the fuel. And if Solaren really thinks it can cut launch costs by the factor of 10 or more needed to make this entire effort viable, then it can't be tripling the cost of the fuel."
The cost of fuel is negligible for today's rockets. Tripling it won't make much difference.
[JR: You miss what I said. They can't use today's rockets. They need to drop launch costs by more than a factor of 10, probably more than 20. That makes fuel costs quite relevant.]
Emissions from rockets vary, but it strikes me as a non-problem in the sense that there are so few launches now that they, too, are negligible, and by the time the launch rate goes up (if it ever does) it will be subject to the same cap-and-trade system everything else will be subject to.
As Leland mentions, there are other lower emission potential means of launch. Right now the market isn't big enough to make them worth pursuing.
For SPS and some other applications, a popular scenario is using material launched from the Moon for the bulk of the structures, processing them in orbit. Needless to say, this requires a big investment to start with in Lunar and orbital infrastructure, so that route is a futuristic one. Launch from the Moon is much easier than from Earth, the Moon has volatiles Leland's looking for (specifically oxygen in lunar regolith for rocket fuel and life support) … but that would all require industrial scale processing. It isn't going to happen with NASA's expensive current Moon plans.
I don't know about the Solaren proposal, but barring some breakthrough I'd expect to see nearer term SPS-like applications before SPS for general power: power relay satellites (Earth-sat-Earth like comsats), Earth-based power beaming, and SPS to supply high-value power (eg: for other space infrastructure, remote locations on Earth, military bases with IED-vulnerable fuel supply routes, disaster relief areas).
I could see some small-scale NASA research and in-space demos of SPS and power beaming concepts at this point. I could also see NASA or DOD (etc) saying "I'll buy X SPS power at Y price" to spur the market. However, NASA probably should concentrate on other things, like more environment monitoring satellites and science, demos of more efficient spacecraft and satellite power-generation and power-using subsystems, more efficient astronaut life-support systems (recycling), monitoring of energy-related processes (winds, grid-effecting space "weather", etc), fuel-efficient aircraft and aircraft operations, etc. NASA could afford a lot more of this if it got rid of the Shuttle and its Ares rocket plans. This would also allow NASA to use commercial rockets and other commercial services, which would be helpful to the economy (and areas like private comsats which are useful for energy-friendly applications like teleconferencing, telemetering, telecommuting, etc).
"Might as well say we're ready to go to the Moon or Mars with private sector financing."
We aren't (we have the technology, but not the market … yet), but we're getting closer. There are numerous private teams going after the Google Lunar X PRIZE to get a rover or some other mobile device on the Lunar surface. These are robotis, but they're working on developing markets besides the Google prize. (They might need NASA as a customer, though … we'll see). Space Adventures has proposed an astronaut trip around the Moon (using Russian government hardware, though).
[JR: You miss what I said. They can't use today's rockets. They need to drop launch costs by more than a factor of 10, probably more than 20. That makes fuel costs quite relevant.]
I agree; at those levels the system would probably be more like an airline where fuel costs would be a big factor in the business. If such a cheap system existed, it would probably get a lot more customers than today's rockets, which would mean that the emissions issue would start to become a real factor for the environment. Hopefully the larger market would enable the launch business to succeed while absorbing the cost of the renewable-based fuel processing.
All this wouldn't help Solaren, though, since it would make it much harder (as if it wasn't hard enough) for their launch vendor to push those launch costs down to the levels Solaren needs.


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A survey by the National Funeral Directors Association notes that people choose to make funeral arrangements ahead of time for two primary reasons: So that their survivors wouldn't have to pay for them or worry about them; and to guarantee the individual's final wishes are taken care of exactly how they want them. Learn about the pre-planning services Holy Cross overs here: http://ow.ly/HuTum
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|Publication number
|Publication type
|Application number
|Publication date
|Jan 16, 1996
|Filing date
|Mar 10, 1994
|Priority date
|Mar 10, 1994
|Fee status
|Also published as
|Publication number
|Martin H. Beck, George F. Rollend, John H. Muszynski
|Original Assignee
|Hoover Universal, Inc.
|Export Citation
|BiBTeX, EndNote, RefMan
|Patent Citations (22), Referenced by (18), Classifications (9), Legal Events (7)
|External Links: USPTO, USPTO Assignment, Espacenet
This invention relates to self-standing containers, preferably bottles, having petaloid bases, for carbonated beverages. Particularly, though not exclusively, the containers comprise biaxially oriented polyester bottles, preferably polyethylene terephthalate (PET), and are manufactured by stretch blow molding.
"Petaloid" is a term of art well understood by those involved in the technology to which this invention relates. However, for the sake of clarity "petaloid" as used herein shall mean "a multi-footed base shape for a self-standing container in which a plurality of feet are disposed in the base portion of the container, somewhat in the general configuration of a plurality of petals" (as in a flower) to provide a stable multi-point self-standing support for the container.
Polyester bottles, for carbonated beverages, having petaloid bases to provide a self-standing ability with a one piece construction are known in the prior art and are commercially available. Such prior art bottles have been constructed of biaxially oriented PET by the known process of blow molding. These prior art one piece bottle constructions have permitted the commercially viable mass production of self-standing beverage containers without the use of a separate base molding utilized in earlier constructions in which the base of the bottle was hemispherical and rendered self-standing by the addition of a separate base unit attached to the hemispherical base by an adhesive. Such two piece constructions do not lend themselves to recycling and were relatively expensive in terms of material and production costs.
Production costs and, more particularly, material costs has led in recent years to substantial research and development in attempts to produce a commercially viable self-standing one piece bottle, for carbonated beverages, which can be produced economically and which performs reliably both in terms of storage and transportation as well as the ability to provide reliable and stable self-standing performance in use.
While this prior art research and development has led to commercially viable products, the viability of these products has been achieved at the expense of the incorporation of additional material to provide the required bottle integrity and stability for commercial use. This additional material was necessitated by the need to design the base of a one-piece bottle which provides a self-standing ability with the consequent elimination of the simple, lightest and most strength effective prior art base form, namely the hemisphere.
Prior art designs, utilizing a hemispherical base, constructed of biaxially oriented PET, such as are found in the commercial market, typically weigh between 46 and 48 grams in the 2 liter size (to this must be added 13 to 16 grams of polyethylene or other plastic material in the separate base which is attached to the hemispherical base to provide standing stability). By comparison, present day commercially available self-standing petaloid base biaxially oriented PET 2 liter bottles weigh from 50 to 56 grams with an average weight of approximately 53.5 grams.
Based on a conservative estimate of 5 billion bottles produced per year and a PET price of $(US)0.7 per pound, a 1 gram increase in the PET content of a bottle will cost approximately $(US)7.7 million per year. Consequently, although the separate polyethylene base is eliminated, the achievement of one-piece self-standing 2 liter PET bottles, meeting the necessary integrity and stability requirements of the industry, as they are currently available has resulted in an increased PET material cost, on the basis of 5 billion bottles per year, of approximately $(US)50 million.
It is an object of the present invention to provide a self-standing petaloid base biaxially oriented polyester bottle for carbonated beverages which is significantly lighter in weight than existing commercially available petaloid base bottle designs without any significant increase in production costs and while meeting industry requirements with respect to integrity and stability during storage, transportation and use.
It is also an object of the present invention to provide improved stress crack resistance of the base.
The light weight biaxially oriented polyester bottle of the present invention achieves weight savings of the polyester material, relative to existing designs of self-standing bottle of similar capacity by the use of unique design concepts relating to the design of its petaloid base. By the use of the unique design features of the present invention, the production of the biaxially oriented PET 2 liter self-standing polyester bottles having a material weight of less than 50 grams, probably as low as 48.0 or even 47.5 grams, while meeting the industry requirements for integrity and stability, is commercially viable. 47.5 grams is within the PET weight range of prior art 2 liter hemispherical base prior art 2 liter biaxially oriented PET carbonated beverage bottles and is 6 grams per bottle lighter than the presently commercially available petaloid base self-standing biaxially oriented PET bottles. The consequence is a material saving, based on 5 billion bottles per year, of approximately $(US)50 million, based on a PET price of $(US)0.7 per pound. This results in a commercially viable 2 liter PET bottle without the PET weight disadvantage of currently commercially available petaloid base self-standing 2 liter bottle designs, while at the same time eliminating the need for the production and use of separate environmentally unsatisfactory polyethylene bases necessitated when hemispherical bases are utilized.
According to the invention there is provided a self standing blow molded polyester container, defining a longitudinal axis, for carbonated beverages having a neck finish integral with and terminating in a neck portion which is integral with and terminates in a side wall portion which is integral with and terminates in a closed base; the neck portion, side wall portion and the base being biaxially oriented; and the base being of a petaloid form defining at least three feet disposed about the longitudinal axis whereby the container is self standing, characterized in that circumferentially adjacent pairs of the feet define, support, and are separated by, relatively stiff valleys each extending substantially radially from a central region of the base, centered on the longitudinal axis, to a relatively deformable open area located radially outwardly of the feet whereby initial internal pressurization of the container will deform the open areas outwardly away from the longitudinal axis to pivot the valleys about their support by the feet thereby to move the central region of the base along the longitudinal axis toward the neck finish.
Also according to the invention there is provided a method of producing a self standing blow molded polyester container, defining a longitudinal axis, for carbonated beverages having a neck finish integral with and terminating in a neck portion which is integral with and terminates in a side wall portion integral with and terminates in a closed base; the neck portion, side wall portion and the base being biaxially oriented; and the base being of a petaloid form defining at least three feet disposed about the longitudinal axis to whereby the container is self standing, characterized by a) providing radially extending relatively rigid valleys between and supported by adjacent pairs of the feet; b) providing relatively deformable areas adjacent radially outer ends of the valleys; c) providing a relatively rigid central region, of the base centered on the longitudinal axis, from which the valleys extend radially; and d) selecting the relative rigidity of the valleys and central region, the support provided by the feet and the relative deformability of the areas whereby upon initial internal pressurization of the container the areas deform outwardly to pivot the valleys about their support by the feet to displace the central region against the internal pressure along the axis toward the neck finish.
The invention will now be described, by way of example, with reference to accompanying drawings, in which:
FIG. 1 is a fragmentary part cross-sectional elevation of a bottle according to the present invention taken on Section Line 1--1 of FIG. 2;
FIG. 2 is an underview of the bottle illustrated in FIG. 1;
FIGS. 3A-7A are diagrammatic inverted (relative to FIG. 1) fragmentary cross-sections taken on Section Lines 3--3 through 7--7 respectively as shown in FIGS. 1 and 2;
FIGS. 3B-7B are diagrammatic representations of centerlines defining the center of material wall thickness illustrated respectively in FIGS. 3A-7A with projection lines illustrating the effective valley widths of the cross-sections illustrated in FIGS. 3A-7A;
FIG. 8 is a fragmentary view taken in the direction 8--8 of FIG. 1;
FIG. 9 is a graphical representation of the deformation of the base of a bottle according to the present invention taken against internal bottle pressure; and
FIG. 10 is a diagrammatic fragmentary cross-section of a valley similar to those illustrated in FIGS. 3A-6A with the addition of a longitudinally extending reinforcement ridge.
Briefly, the base of the preferred form of container of the present invention comprises a petaloid design for a bottle which has a plurality of at least three (preferably five) feet evenly disposed around the longitudinal axis of the bottle and projecting from a hemispherical base form of the bottle to provide a stable self-standing support for the bottle. Between each adjacent pair of feet is a radially extending valley, the valley floor of which is preferably curved in cross-section (in one form a secondary fold or radially extending ridge stiffens and reinforces the valley floor). The valley width decreases with increasing radial distance from the bottles longitudinal axis so that the walls of the valley tend toward a point of convergence radially outside the bottle's diameter. The valley floor generally follows the hemispherical base shape and opens to an extended portion of that base shape lying radially outwardly of the feet.
The shape, dimensions and material thickness etc. of the valley and extended portion are chosen so that initial pressurization of the bottle tends to expand and deform the extended portions outwardly with a resulting raising of the center of the base, at the longitudinal axis, away from the support surface. Further pressurization will reverse this and the center of the base may return at least to its unpressurized location. This action reduces the downward destabilizing deformation of the center of the base as compared with existing petaloid base designs and permits a lighter construction while still meeting industry performance requirements.
Referring first to FIGS. 1 and 2, a one piece self-standing biaxially oriented PET two liter bottle 1, of circular horizontal cross-section, comprises a neck finish 2 connected to a neck transition portion 3 of the bottle by way of a neck support ring 4. The neck transition portion 3 connects by way of an upper portion 5 of the bottle to a substantially cylindrical side wall portion 6 which terminates at its lower end in a closed base 7, the underlying shape of which is hemispherical. The bottle 1 defines a longitudinal axis 8.
Projecting downwardly from the hemispherical form of the base are five hollow feet 9 which together form a petaloid foot formation with the feet symmetrically and evenly disposed about the longitudinal axis 8 to provide the stable support for the bottle necessary to provide its self-standing ability. The lowest extensions of the feet 9 terminate in bottle support pads 10. Each foot 9 comprises sloping walls 11 extending from its pad 10 to its junction with the underlying hemispherical formation (reference numbers for sloping walls 11 are illustrated in FIG. 2 only with respect to one of the feet although all of the feet are identical).
Radially extending valleys 12 are disposed between adjacent pairs of feet 9. These valleys 12 each include a valley floor which substantially follows the surface curvature of the underlying hemispherical shape of the base 7 and terminates at and open into an extended portion 13 (see FIG. 8). Although shown by solid lines (FIG. 2) for simplicity at the junctions between the sloping walls 11 and the valleys 12 and pads 10, the intersection of these elements are curved in cross-section to provide smooth transitions and structural rigidity of the valleys along their length.
All of the valleys 12 are substantially identical and each valley converges in effective width toward a point of convergence 14 lying outside of the outer diameter of the bottle 1 (FIG. 2).
The central area of the base 7, through which extends the axis 8 is connected to each pad 10 by a substantially flat ridge path 15 joined on either side to portions of the sloping walls 11.
The bottle illustrated includes a small annular lip 16 which is primarily present for aesthetic purposes and for label alignment during production. This lip lies adjacent the transition from the sidewall 6 to the base 7.
With particular reference to FIG. 1 it will be noted that although the bottle is illustrated in cross-sectional form and although the material of the bottle will usually be substantially transparent, details of the interior of the bottle, lying beyond a cross-section taken are omitted for the sake of clarity in the illustration of the invention.
Now referring to FIGS. 3A/B through 7A/B the structure of one of the five identical valleys will be described. In each of the five cross-sections the A designation indicates a fragmentary cross-section of the valley concerned while the "B" designation represents the center of thickness of material, shown in the associated "A" designation cross-section, with projection lines illustrating the effective structural valley width of that associated cross-section.
FIG. 3A, taken on Section Lines 3--3 of FIGS. 1 and 2, is the valley cross-section closest to the longitudinal axis 8 of the bottle and illustrates the increased thickness of the material of the bottle in the region of the longitudinal axis 8 and the portion of the valley most closely adjacent that axis. The variation of the base thickness along the length of the valley is best illustrated to the right of the longitudinal axis 8 in FIG. 1. The increased thickness in the central area of the base of the bottle is required to prevent inversion (excess downward deformation) of the central area of the base in use (storage, transportation and beverage consumption related activities) when pressurized by a carbonated beverage. FIGS. 4A, 5A and 6A illustrate fragmentary cross-sections of the valley at the Section Lines 4--4, 5--5 and 6--6 of FIGS. 1 and 2 and show the decreasing depth and width of the illustrated valley the radial distance, from the longitudinal axis 8 increases. The construction lines 17 of FIGS. 3B-6B represent an extension of the sloping wall 11 to the base of the valley in order to illustrate the effective width of the valley as it decreases with an increase in radial distance from the axis 8. This reduction in width is shown by the dimension X, X-1n, X-2n and X-3n in FIGS. 3B, 4B, 5B and 6B, respectively, with X being the respective width of the valley at the cross-section 3--3 and n being a number representative a decrease in effective valley width from illustrated section to illustrated section.
FIGS. 7A and B illustrate a cross-section of the base of the bottle in the extended portion 13 radially outside of the cross-section illustrated in 6A and into which the valley opens. The extended portion 13 represents an area of the base adjacent its greatest diameter and close to the transition from the base to the sidewall portion The extended portion extends around a substantial portion of the circumference of the base to define a relatively deformable region of the base adjacent and in communication with the radially outer end of each valley. In addition to the figures already discussed, reference should be made to FIG. 8 which illustrates the relationship between each valley and its associated extended portion of the base.
The relatively deformable extended portions 13 and the relatively rigid valleys 12 supported by the relatively rigid feet 9 together serve to allow the construction of the present invention to perform in a manner meeting the integrity and stability requirements of the industry while enjoying a substantial reduction in weight (from an average of 531/2 grams to approximately 471/2 grams for a two liter bottle) by comparison with existing petaloid based self-standing biaxially oriented PET two liter bottles for carbonated beverages.
The described elements of the present invention function to provide the superior performance as follows. Upon initial pressurization of the bottle,the extended portions are deformed outwardly with a resulting application of force to pivot the substantially rigid levers, provided by the valleys 11, about a substantially rigid pivotal location provided by the substantially rigid feet 9 with the consequent uplifting of the central region of the base of the bottle at the location of the longitudinal axis 8, namely the area of the base of the bottle which is most prone to inversion (excessive downward deformation) upon the application of internal pressure to the bottle. This effect increases during an initial increase in internal pressure in the bottle until limitations in the deformation of the extended portions 13 and of the rigidity of the valleys 11 and feet 9 result in the internal pressure in the bottle overcoming the upward bias of the central portion of the base and reversing that upward deformation until the central portion adjacent the location of the longitudinal axis 8 is deformed downwardly to and past its location when zero pressure is applied internally to the bottle. Thus, a substantial internal pressure is supported in the bottle before the central portion of the base begins to deform downwardly, below its zero pressure location, thereby facilitating the required performance of the bottle with respect to integrity and stability at a substantially lighter material weight than that of prior art petaloid bottles in which the deformation of the corresponding central region of the base in a downward direction commences immediately upon the application of pressure to the inside of the bottle.
With reference to FIG. 9 there is illustrated a graphical representation of the deformation characteristics of the central portion of the base of a bottle a) according to the present invention (solid line 18) with b) a typical prior art petaloid base bottle (chain dashed line 19) weighing approximately 6 grams more than the bottle of the present invention and c) the projected deformation (dashed line 20) of a petaloid based two liter biaxially oriented PET bottle similar to that already existing in the prior art but with a weight reduction in the base area of the bottle corresponding to the weight reduction achieved by the present invention but without the innovative design characteristics of the base portion of the present invention. As Can be seen, the central base region of the bottle of the present invention is deformed upwardly with an initial increase in internal pressure of the bottle from zero and then, with a further increase in internal pressure, returns to its initial zero pressure position and passes through that position to a downward deformation as internal pressure is further increased. The heavier existing prior art bottle represented by the deformation curve 19, shows a continuous downward deformation of the central region of the base from the initial application of internal pressure. Both the bottle of the present invention and the existing heavier prior art bottle represented by the deformation curves 18 and 19 are able to meet industry standards for integrity and stability. However, the projected curve 20 of a lightened prior art bottle otherwise similar to the bottle resulting in the performance curve 19, shows a markedly increased deformation of the base region of the bottle which will result in premature failure or excessive deformation which will not permit that bottle to meet the aforementioned industry standards. Deformation curves 18 and 19 are diagrammatic representations of actual test results while the deformation curve 20 is an illustration of the projected deformation of the bottle concerned extrapolated from the known deformation characteristics of the known prior art bottles and stress analysis of such a bottle with the decreased base weight involved.
FIG. 10 shows variation on the embodiment illustrated in FIGS. 1-8. In this embodiment, each of the valleys 12 includes in its floor a ridge or secondary fold 21 extending along its length to additionally stiffen the valley. The ridge 21 may extend for substantially the entire length of the valley from the cross-section illustrated in FIG. 3A to the cross-section illustrated in FIG. 6A.
|Cited Patent
|Filing date
|Publication date
|Dec 15, 1971
|Sep 18, 1973
|Owens Illinois Inc
|Pressure resistant plastic container
|Sep 6, 1972
|Sep 24, 1974
|Phillips Petroleum Co
|Molecularly oriented hollow article, such as a bottle
|May 23, 1977
|Aug 22, 1978
|The Continental Group, Inc.
|Ribbed bottom structure for plastic container
|Aug 13, 1979
|Jan 27, 1981
|Sewell Plastics, Inc.
|Bottom structure for plastic container for pressurized fluids
|Aug 9, 1979
|Mar 10, 1981
|Yoshino Kogyosho Co., Ltd.
|Plastic pressure bottle
|Jun 8, 1982
|Sep 13, 1983
|The Continental Group, Inc.
|Plastic container with hollow internal rib reinforced bottom and method of forming the same
|Nov 30, 1979
|Jun 25, 1985
|The Continental Group, Inc.
|Plastic container with internal rib reinforced bottom
|Mar 12, 1986
|Nov 22, 1988
|Continental Pet Technologies, Inc.
|Plastic bottle base reinforcement
|Dec 24, 1987
|Sep 5, 1989
|Continental Pet Technologies, Inc.
|Hot fill container
|Jan 23, 1989
|Sep 12, 1989
|Hoover Universal, Inc.
|Blow molded one-piece bottle
|Jul 15, 1988
|Sep 19, 1989
|Hoover Universal, Inc.
|Blow molded bottle with improved self supporting base
|Jul 15, 1988
|Jan 9, 1990
|Hoover Universal, Inc.
|Concentric ribbed preform and bottle made from same
|Apr 11, 1989
|May 22, 1990
|Continental Pet Technologies, Inc.
|Preform and method of forming container therefrom
|Jan 10, 1990
|Dec 18, 1990
|North American Container, Inc.
|Plastic container for pressurized fluids
|Oct 4, 1990
|Jun 18, 1991
|Sewell Plastics, Inc.
|Wide stance footed bottle
|Dec 11, 1990
|Dec 10, 1991
|Hoover Universal, Inc.
|Blow molded PET container with ribbed base structure
|Apr 2, 1987
|Nov 3, 1992
|Continental Pet Technologies, Inc.
|Reinforced container base and method of forming same
|Jun 9, 1992
|Apr 27, 1993
|Constar Plastics, Inc.
|Footed container
|Jul 16, 1992
|Feb 22, 1994
|Plastipak Packaging, Inc.
|Plastic blow molded freestanding container
|Jun 8, 1992
|Jun 14, 1994
|Yuan Fang Limited
|Base configuration for biaxial stretched blow molded pet containers
|Mar 26, 1993
|Jun 27, 1995
|Continental Pet Technologies, Inc.
|Freestanding container with improved combination of properties
|Title not available
|Citing Patent
|Filing date
|Publication date
|Jul 7, 1997
|Dec 22, 1998
|Dtl Monofoot Limited Partnership
|Base design for one piece self-standing blow molded plastic containers
|Jul 11, 1997
|Jul 27, 1999
|Pepsico, Inc.
|Pressured thermoplastic beverage containing bottle with finger gripping formations
|Jul 10, 1998
|Nov 23, 1999
|Crown Cork & Seal Technologies Corporation
|Footed container and base therefor
|May 1, 1998
|May 16, 2000
|Crown Cork & Seal Technologies Corporation
|Hot fill plastic container having spaced apart arched ribs
|Sep 10, 1998
|Sep 5, 2000
|Bcb Usa, Inc.
|Container with base having cylindrical legs with circular feet
|Nov 22, 1999
|Apr 10, 2001
|Crown Cork & Seal Technologies Corporation
|Footed container and base therefor
|Aug 26, 1998
|Oct 2, 2001
|Crown Cork & Seal Technologies Corporation
|Mold used to form a footed container and base therefor
|Dec 7, 1999
|Oct 30, 2001
|Rehrig Pacific Company
|Storage container
|Nov 20, 1997
|Dec 4, 2001
|General Mills, Inc.
|Plastic container for food products
|May 15, 2000
|Feb 19, 2002
|Crown Cork & Seal Technologies Corporation
|Hot fill plastic container having spaced apart arched ribs
|Feb 1, 2002
|Dec 23, 2003
|Pepsico Inc.
|Plastic container having an outwardly bulged portion
|Dec 7, 2006
|Nov 8, 2011
|Sidel Participations
|Drawing-blowing method and device for molding thermoplastic material containers, in particular bottles, with petaloid bottoms
|Jan 21, 2010
|May 6, 2014
|General Mills, Inc.
|Thermoformed container assembly for food products
|Dec 7, 2006
|Jun 14, 2007
|Sidel Participations
|Drawing-blowing method and device for molding thermoplastic material containers, in particular bottles, with petaloid bottoms
|Apr 23, 2008
|Oct 23, 2008
|Esmeralda Reyes Rossi
|Self Righting Baby Bottle
|Jul 5, 2012
|Krones Ag
|Container of a thermoplastic material
|Sep 10, 1999
|Mar 16, 2000
|Bcb Usa Corp.
|Container with base having cylindrical legs with circular feet
|Dec 4, 2000
|Jun 14, 2001
|Rehrig Pacific Company
|Storage container
|U.S. Classification
|215/375, 264/532, 220/609, 264/523, 215/381
|International Classification
|Cooperative Classification
|European Classification
|Apr 4, 1994
Owner name: DTL TECHNOLOGY LIMITED PARTNERSHIP, NEW HAMPSHIRE
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:BECK, MARTIN H.;ROLLEND, GEORGE F.;MUSZYNSKI, JOHN H.;REEL/FRAME:006924/0764;SIGNING DATES FROM 19940308 TO 19940309
|Aug 29, 1994
Owner name: HOOVER UNIVERSAL, INC., MICHIGAN
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:DTL TECHNOLOGY LIMITED PARTNERSHIP;REEL/FRAME:007109/0766
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Owner name: SCHMALBACH-LUBECA AG, GERMANY
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:JOHNSON CONTROLS, INC.;HOOVER UNIVERSAL, INC.;REEL/FRAME:008454/0072
Effective date: 19970228
|May 28, 1999
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|Jul 8, 2003
|Fee payment
Year of fee payment: 8
|Aug 1, 2003
Owner name: AMCOR LIMITED, AUSTRALIA
Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNOR:SCHMALBACH-LUBECA AG;REEL/FRAME:014294/0971
Effective date: 20021208
|Jul 13, 2007
|Fee payment
Year of fee payment: 12


text:
At Whitewater Ski Resort, we like to keep things simple, affordable and accessible for everyone! We are pleased to offer a variety of rates to suit each age group. Don't miss the Early Bird Rates, which end on September 30, 2015 for our Season Passes.
For our Alpine Day Rates click HERE. Whitewater Ski Resort is happy to offer half-day rates beginning at *12:30pm for those who prefer to hit the slopes for the later part of the day. As always, our Powder Plus Card is a great deal when skiing multiple days.
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horse training child/adult lessons hunters/jumpers/equatation youth western lessons
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sales jobs near Salisbury, MD
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Capital Blue Cross Medical & Dental Plan benefits. Solar Sales Consultant Overview*. Do you have a consultative sales approach that gains the trust of your...
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A college degree or equivalent sales experience. We will consider all qualified candidates regardless of race, color, national origin, sex, age, marital status,...


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Date: 1886
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the rubber gasket at the breaks was leaking. fixed that but now there is almost no pressure in the line and there is an air hissing noise comming from the pedal. bled break line and got better but not by much. don't know what to check next.
break peddal has no pressure & hissing at peddal. bled system. now what? on 1994 Saturn SL2
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3 answers
maybe your brake booster Is bad
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1M Series Halogen Waterproof Alkaline Spotlight by Stanley®. This affordable, waterproof halogen hand-held spotlight uses a 13W quartz-halogen automotive-style H3 bulb and the traditional power for flashlights – C-cell alkaline batteries, 8 of them in all. It's waterproof, capable of total immersion up to six feet underwater and for as long as an hour without getting waterlogged.
- Halogen Bulb: 12V, H3 series, 13 watt
- Power Source: 12V DC power cord (included)
- Or 8 "C" type alkaline batteries (not included)
- Sturdy plastic housing
- O-rings on power port, rubber bezel and trigger switch keep it dry inside
Pumping out plenty of light, this waterproof flashlight is perfect for boaters, fishermen and hunters, or anyone who needs to see where they're going in the dark and the rain. Not only will it run on its internal flashlight batteries, it comes with a 12V power cable that connects to your vehicle's power plug, whether it's a car, truck, boat, ATV or motorcycle. When you plug this cord into the spotlight, it switches over to auxiliary power automatically, saving your batteries for another time. If you need to exit the vehicle, just unplug the cord from the back of the light, and go back to the internal batteries.
The Stanley Works was founded in 1843 in New Britain, Connecticut by Frederick T. Stanley to manufacture door hinges and bolts.. A few years later, in 1857, Frederick's cousin, Henry founded The Stanley Rule and Level Company. In 1910, S. Duncan Black and Alonzo G. Decker started their shop in Baltimore, Maryland, and were awarded a patent 6 years later for the world's first portable power tool. In 2010 Stanley and Black & Decker combined to form Stanley Black & Decker, to deliver the tools and solutions that industrial companies, professionals, and consumers count on to be successful when it really matters.