A method for the preparation of Mycobacteria from any liquid, semi solid or exotic source is described. The extracted Mycobacterial sample is suitable for detection by culture and amplification.

A. Mycobacteria and Disease
Mycobacterium tuberculosis (MTB) is the causative agent of the most common infectious disease in the world today, tuberculosis (TB). The World Health Organization (WHO) reported that 1.7 billion people (or approximately one-third of the world's population) are currently, or have been, infected by tuberculosis (Kochi, A. Tubercle 72:1-6 (1991)). The incidence of MTB infection is occurring at an increasing rate with 8 million new cases worldwide in 1991 (Sudre, P. et al., Bull. W.H.O. 70:149-159 (1992)), and the WHO estimates that 88.2 million people will contract TB during the 1990's and approximately 3 million people will die annually during this time period (Morbidity and Mortality Weekly Report 42 (No. 49):961-964 (1993)). In the United States the Centers for Disease Control and Prevention (CDC) recorded 26,673 cases in 1992 (Morbidity and Mortality Weekly Report 42:696-703 (1993)), and it is estimated that 10 to 15 million people in the U.S. have latent infections (Morbidity and Mortality Weekly Report 39(RR-8):9-12 (1990)).
The importance of Mycobacteria of the MAC complex, (primarily M. avium and M. intracellulare), as human pathogens was recently reviewed by Inderlied, C. B. et al., Clin. Microbiol. Rev. 6:266-310 (1993). MAC complex infections have been on the rise owing to their occurrence as opportunistic pathogens in AIDS patients. Approximately 43% of AIDS patients, with advanced stages of the disease, present with disseminated MAC infections (Nightingale et al., Jour. Infect. Dis. 165:1082-1085 (1992)). The WHO estimates that today approximately 3 million people have developed AIDS, approximately 15 million have been infected with the human immunodeficiency virus (HIV), and by the year 2000 the number infected could climb to approximately 40 million (World Health Organization (document WHO/GPA/CNP/EVA/93.1) Global Programme on AIDS (1993)). In addition to AIDS related infections, M. paratuberculosis, a subspecies of M. avium (Thorel, M. F. et al., Int. J. Syst. Bacteriol. 40:254-260 (1990)), is thought to be associated with Crohn's disease, an inflammatory disease of the bowel (Chiodini, R. J. Clin. Micro. Rev. 2:90-117 (1989)).
Mycobacterial infections are also a problem in animals. M. paratuberculosis also causes bowel inflammations in ruminants (Thoen, C. O. et al., Rev. Infect. Dis. 3:960-972 (1981)). This is more commonly known as Johne's disease. Cattle that test positive for Johne's are culled and destroyed. In the state of Wisconsin, where approximately one-third of the herds are infected (Collins, M. T., Hoard's Dairyman Feb 10:113 (1991)), the financial impact was estimated at $52 million in 1983 (Arnoldi, J. M. et al., Proceedings, 3rd Int. Symp. World Assoc. Vet. Lab. Diag. 2:493-494 (1983)). The incidence amongst herds nationwide typically ranges between 3% and 18% (Merkal, R. S. et al., J. Am. Vet. Med. Assoc. 190:676-680 (1987)). The financial impact of this one disease on the dairy industry exceeds $1.5 billion annually (Whitlock, R. Proceedings of the Third International Colloquium on Paratuberculosis, pp.514-522 (1991); Whitlock, R. et al., Proceedings of the 89th Annual Meeting of the United States Animal Health Association, pp.484-490 (1985)).
In addition to the organisms discussed above, a wide variety of Mycobacteria are also considered human pathogens, including Mycobacterium leprae, Mycobacterium kansasii, Mycobacterium marinum, Mycobacterium fortuitum complex, and many others. Wayne, L. G. et al., Clin. Micro. Rev. 5:1-25 (1992) review the diversity of infections associated with this genus of microorganism. However, the magnitude and impact of these infections is not on the same scale as MTB complex and MAC infections. For example, leprosy is probably the most common within this category: there are an estimated 5.5 million cases of Mycobacterium leprae worldwide (Nordeen, S. K. et al., Int. J. Lepr. 63:282-287 (1993)). Taken as a whole, this group of organisms exacts a tremendous social cost.
B. Culture and Detection of Mycobacteria
The contemporary protocol(s) for the laboratory diagnosis of Mycobacterial infections are relatively slow. Extended incubations are required owing to the innate slow growth rate of these bacteria. Owing to this lengthy time to diagnosis, individuals suspected of infection are quarantined, or else pose significant risk to society in general.
In addition, laboratory confirmation of the diagnosis of Mycobacterial infections requires several cultures per patient sample. Each sample must be incubated up to eight weeks (sixteen weeks for M. paratuberculosis) before the sample can be reported negative. The need for multiple cultures of each suspected sample is due in part to the intermittent shedding of detectable numbers of Mycobacteria, and the loss of infectious organisms due to the harsh chemical decontamination used to inactivate saprophytic microorganisms. These procedures are inefficient and often kill the Mycobacteria they are attempting to extract. For example, processing by the recommended N-acetyl-L-cysteine-NaOH (NALC/NaOH) procedure (Kent, P. T. et al., "Public Health Mycobacteriology," in A Guide for the Level III Laboratory, U.S. Department of Health and Human Service, Centers for Disease Control, (1985) pp. 31-46) is known to kill 28%-33% of the existing Mycobacteria (Krasnow, I. et al., Am. J. Clin. Path. 45:352-355 (1966); Kubica, G. P. W. et al., Am. Rev. Resir. Dis. 87:775-779 (1963)). The advent of contemporary probe assays (Gonzalez, R. et al., Diag. Microbiol. Infect. Dis. 8:69-78 (1987)) that complement culture techniques has improved the time to diagnosis; however, there still exists room for significant improvement.
The combination of social importance and reliance on culture methods reveals a critical need for a Mycobacterial testing protocol that reduces turn around time and increases sensitivity. The isothermal scheme being commercialized by Gen-Probe, Inc. (San Diego, Calif.: Jonas, V. et al., J. Clin. Micro. 31:2410-2416 (1993)) and the polymerase chain reaction (PCR) both have the potential for single molecule detection (Higuchi et al., Nature (London) 332:543-546 (1988)). Furthermore, amplification and detection can be performed in approximately eight hours and the reagents do not add a significant cost. If available, an amplification assay could greatly enhance the speed and sensitivity of detection, and reduce the cost of Mycobacterial diagnosis (De Cresce, R. P. et al., Med. Laboratory Obs. 25:28-33 (1993)). The rapidity with which these technologies could potentially diagnose Mycobacterial infections would have a tremendous financial impact on society.
However, as described herein, researchers have encountered a plethora of problems in an effort to adapt these technologies, such as PCR amplification, to the detection of Mycobacteria. Especially it has not been possible to develop a protocol for the preparation of a sample for analysis in a manner that will (a) ensure amplification assay detection of a true positive result and also (b) not give false negative results. The variability encountered by researchers is exemplified by the study of Noordhoek, G. T. et al., J. Clin. Micro. 32:277-284 (1994). These authors describe a blind study in which seven laboratories participated. All laboratories used the same amplification system, but different processing and detection methodologies. The original summary of these results (Noordhoek, G. T. et al., N. Eng. J. Med. 329:2036 (1993) concisely shows that at low copy numbers (1000 copies), the correlation varied from 2% to 90%, with the average being 54%. As a result of these problems, there is still no available FDA approved TB-amplification kit.
C. Methods of Processing Mycobacterium Samples
A review of the scientific literature on Mycobacteria-nucleic acids amplification references relating to system design, sample processing techniques and clinical studies reveals the highly variable results that have precluded FDA approval of a TB-amplification kit.
Two different amplification schemes have been used; there are numerous PCR system designs and many clinical studies that focus on Mycobacterial infections, the vast majority on MTB. Typically, samples are processed for culture first and then subjected to amplification. Therefore, sample preparation for amplification can be viewed, in most cases, as an extension of the culture processing protocol.
There are several reasons the field has evolved in this manner. First, obtaining clinical specimens is difficult. Individuals diagnosed with MTB are invariably started on drug therapy upon diagnosis. Second, processing MTB specimens requires specialized containment facilities and appropriately trained technicians. Third, it is the easiest way to obtain "culture correlation" results; that is, a correlation of amplification positive and negative results with those that were positive or negative in culture. Consequently, researchers have typically processed the sample for culture and then used protocols that "further" process the sediment for amplification. In this way, actual clinical specimens can be used, containment is not breached, work flow is not interrupted, patient care is not compromised and correlation to contemporary protocols is feasible. This "further" processing referred to above has involved a wide variety of sample preparation and cell lysis techniques.
Table 1 summarizes 35 publications, using samples derived from 17 different countries, evaluating the performance of amplification technologies in the clinical laboratory. The works in Table 1 are presented chronologically. Every effort has been made to accurately represent the original publication. There were, however, ambiguities in interpretation in some instances, and distinctive features of several papers. Clarifications are highlighted in the footnotes that follow.
Correlation of nucleic acid amplification results with culture results was chosen as the basis to compare the studies shown in Table 1. Using this perspective, this analysis reveals a conundrum: according to the methodologies outlined in Table 1, the sample subjected to amplification is derived, in the vast majority of instances, from the "button" used to seed the culture. Given the sensitivity of amplification relative to culture, a culture positive/amplification negative (e.g., false negative amplification) does not make intuitive sense. Several authors present "corrected correlations" (see Footnote C in Table 1). For example, if several false negatives were obtained, and the result could be resolved by further purification of the target DNA, dilution of inhibitors, multiple amplifications of the same sample, multiple amplifications of different samples from the same patient, or reamplification of the amplified specimen; the corrected results were presented.
This introduces an interesting dilemma. Jackson, J. B. et al., J. Clin. Micro. 31:3123-3128 (1993)) have successfully implemented an HIV-PCR quality assurance panel involving 11 laboratories. The reported sensitivity suggests that all laboratories have the routine capability of identifying 2 copies of the HIV genome in a background of 10.sup.6 human cells. The studies in Table 1 that are reviewed herein strongly suggest that the sensitivity of TB-amplification technologies is similar. What is apparent from the discussion of the studies in Table 1 is that, while the sensitivity of these amplification technologies for detection of Mycobacteria is expected to be orders of magnitude greater than that of either culture or smear, there are consistent aberrations to this expectation. While some of the anomalies are due to inhibition, and are, therefore, easily explained, many of the irregularities are "unexplained." What will become apparent is that, even if an internal control is used to detect inhibitors, these unexplained aberrations are reasonably common, and as such pose a significant obstacle to validation of amplification technologies for the detection of Mycobacteria in the clinical laboratory. For example, while positives, or suspected positives, may be rechecked, typically negatives are not. Since false negatives will occur with regularity, the laboratory will have no way of knowing which samples are truly false negative and which need to be "resolved." Consequently, patient care will be compromised as the diagnosis will incorrectly appear negative. Therefore, for the purpose of discussion of the published art herein, the art's uncorrected data was used. These aberrations--the false negatives--are the focus of the remaining discussion.
TABLE 1 __________________________________________________________________________ Art Reported Correlation of Culture Results with Nucleic Acids Amplificati on Results Author(s) .sup.A Processing Protocol .sup.B Total: Cult .sym./Amp .sym. .sup.C Correl. __________________________________________________________________________ Brisson-Noel et al., Lancet ii:1069-1071 SDS.sub.P => NaOH/SDS/95.degree./15' => Org/ppt 35: 13/15 100% (1989) Shankar et al., Lancet 335:423 (1990) NALC => NaOH/NaCl/SDS/95.degree./1 5' => Org/ppt .sup.D 23: 8/10 100% Hermans et al., J. Clin. Micro. 28:1204-1213 => (NaOH/NaHPO.sub.4).sub.P => TE).sub.W => Lz/SDS/PrK => Org/ppt .sup.E 17: 7/11 100% (1990) Pao et al., J. Clin. Micro. 28:1877-1880 (1990) OxAc => NaCl/EDTA => Lz => Phenol/SDS => ppt 284: 49/118 (100%).sup.F .dagger-dbl.Sjobr ing et al., J. Clin. Micro. 28:2200-2204 DTT => TE/95.degree. => Sonic .sup.g 7: 4/3 75% (1990) DEWit et al., J. Clin. Micro. 28:2437-2441 => PEG => 70.degree./30' => Phenol/SDS => Org/PEG-ppt .sup.H 26: 14/14 100% (1990) Thierry et al., J. Clin. Micro. 28:2668-2673 (=>) NaOH/SDS/95.degree./15 ' => Org/ppt .sup.I 75: 30/35 100% (1990) .dagger..dagger-dbl.Pierre et al., J. Clin. Micro. 29:712-717 (a) SDS.sub.P => NaOH/SDS/95.degree ./15' => Org/ppt .sup.J 82: 24/23 (79.2%).sup.K (1991) (b) SDS.sub.P => (PBS).sub.W => PCR.sub.Buf /PrK/NonI => 95.degree. C./10' Del Portillo et al., J. Clin. Micro. => (Water/95.degree./10').sub.P => Lz => SDS/PrK => Org/ppt .sup.L 30: 11/18 100% 29:2163-2168 (1991) .dagger..dagger-dbl..sctn.Brisson-Noel et al., Lancet 338:364-366 SDS.sub.P => NaOH(SDS/95.degree ./15' => Org/ppt .sup.M 446: 141/110 (89.4%).su p.N (1991) .dagger-dbl.Sritharan et al., Mol Cell. Probes 5:385-395 NALC.sub.P => (TEX).sub.W => TEX/95.degree./30' .sup.O 96: 74/88 (95.9%).sup.P (1991) Eisenach et al., Am. Rev. Resp. Dis. NALC.sub.P => Lz => NaOH(SDS/95.deg ree./5' => GuSCN/Si 162: 48/53 97.7% 144:1160-1162 (1991) Manjunath et al., Tubercle 72:21-27 (1991) NALC => NaOH/SDS/95.degree./1 5' => Org/ppt 117: 17/31 100% .paragraph.Cousin s et al., J. Clin. Micro. 30:255-258 NALC.sub.P => (Water).sub.W => Lz => SDS/PrK => TMA/Org/ppt 177: 64/92 98.4% (1992) .dagger.VAN DER Giessen et al., J. Clin. Micro. (a) NaOH.sub.S => Phenol/CHCl.sub.3 => Silica => H.sub.2 O .sup.Q 87: 30/5 & 1 16%, 3% 30:1216-1219 (1992) (b) NaOH.sub.S => Phenol/CHCl.sub.3 => Silica => H.sub.2 O 87: 30/7 & 1 23%, 3% (c) IDEXX.sub.BU F => Column => Pellet => NaOH => 120.degree./10' 87: 30/4 & 4 13%, 13% .dagger..dagger-dbl..paragraph.Buck et al., J. Clin. Micro. 30:1331-1334 NALC.sub.P => 2x(Water).sub.W => Sonic => 95.degree./10' .sup.R 43: 25/24 92.0% (1992) .dagger..dagger-dbl..sctn.Victor et al., J. Clin. Micro. 30:1514-1517 (a) NALC => PCR.sub.Buf => 95.degree./10' .sup.S 155: 131/100 76.3% (1992) (b) NALC => Sucrose => (PBS).sub.W => PBS 155: 131/131 98.5% .dagger.Soini et al., J. Clin. Micro. 30:2025-2028 NaOH/SDS.sub.P => NaOH/SDS/95.degree ./15' => Org/ppt 127: 34/25 55.9% (1992) Altamirano et al., J. Clin. Micro. 30:2173-2176 NALC.sub.P => Lz/SDS => perCl => Org/ppt 200: 44/43 97.7% (1992) .dagger.Fauville-Dufaux et al., Eur. J. Micro. Inf. Dis. SDS or TriPO.sub.4 => NaOH/SDS/95.degree ./15' Org/ppt .sup.T 206: 92/84 (91.3%).sup.U 11:797-803 (1992) .dagger..dagger-dbl.Kolk et al., J. Clin. Micro. 30:2567-2575 NALC.sub.P => (TX).sub.W => TX/PrK/60.degree./ 18 hrs => 95.degree./15' 227: 45/81 95.6% (1992) .dagger..dagger-dbl..sctn..paragraph.Shawar et al., J. Clin. Micro. 31:61-65 NALC.sub. P => (TEX).sub.W => TEX/95.degree./ 30' 384: 71/75 78.9% (1993) .sctn..paragraph.Wilson et al., J. Clin. Micro. 31:776-782 (a) NaOH => (PBS).sub.W => PBS/80.degree./20' => GuSCN/Si .sup.V 171: 27/19 (66.7%).sup.W (1993) (b) NaOH => (PBS).sub.W => PBS/80.degree./20' => CHCl.sub.3 171: 27/26 (88.9%) .dagger.Folgueira et al., J. Clin. Micro. 31:1019-1021 NALC.sub.P => PCR.sub.Buf PrK/NonI => 95.degree./10' .sup.X 75: 71/75 (90.7%).sup.Y (1993) Kocagoz et al., J. Clin. Micro. 31:1435-1438 NALC.sub.P => 3x(TE).sub.W => TE/95.degree./1 0' .sup.Z 78: 29/36 (100%).sup.. alpha. (1993) .dagger..dagger-dbl..paragraph.Forbes et al., J. Clin. Micro. 31:1688-16 94 NaOH => PBS/NonI => 95.degree./10' => Sonic .sup..beta. 173: 31/25 (80.6%).sup..gamma . (1993) 727: 80/75 (83.8%) .dagger..paragraph.Nolte et al., J. Clin. Micro. 31:1777-1782 NALC.sub.P => TEX => 95.degree./30' 313: 123/113 91.1% (1993) .dagger..dagger-dbl.Irula et al., J. Clin. Micro. 31:1811-1814 => Fic/Hyp/PBMC => (TEX).sub.W => ChX/56.degree./30' => 95.degree./30' .sup..delta. 243: 15/44 80.0% (1993) .dagger..dagger-dbl..sctn..paragraph.Clarridge et al., J. Clin. Micro. NALC.sub.P => (TEX).sub.W => TEX/95.degree./30' 1,166: 218/192 (83.0%).sup..epsil on. 31:2049-2056 (1993) Miyazaki et al., J. Clin. Micro. 31:2228-2232 DTT => Lz => PrK/SDS => Org/ppt 417: 56/92 (96.4%).sup. .zeta. (1993) .dagger..dagger-dbl..sctn.Jonas et al., J. Clin. Micro. 31:2410-2416 NALC => HCl => Gen.sub.Buf => Sonic => 95.degree ./15' .sup..eta. 758: 119/116 (79.8%).sup..theta . (1993) .dagger-dbl..sctn..paragraph.Abe et al., J. Clin. Micro. 31:3270-3274 (a) Gen: NALC => PBS/HCl => Gen.sub.Buf => Sonic => 95.degree ./15' .sup..iota. 135: 32/34 (90.6%).sup..kappa . (1993) (b) PCR: NALC => TEX/GuSCN/Si 135: 32/32 (81.2%) .dagger..dagger-dbl..paragraph.Miller et al., J. Clin. Micro. 32:393-397 (a) Gen: NALC => HCl => Gen.sub.Buf => Sonic => 95.degree./15' .sup. 750: 142/131 (83.9%).su p..mu. (1994) (b) PCR: (NALC).sub.P => PCR.sub.Buf => Sonic => 95.degree./15' 156: 142/122 (78.2%) .dagger..sctn.Pfyffer et al., J. Clin. Micro. 32:918-923 (a) NALC => HCl => Gen.sub.Buf => Sonic => 95.degree./15' .sup..nu. 515: 42/57 (92.9%).sup. .xi. (1994) (b) SDS => HCl => Gen.sub.Buf => Sonic => 95.degree./15' 423: 36/50 (97.2%) .dagger-dbl.Bodme r et al., J. Clin. Micro. 32:1483-1487 SDS => HCl => Gen.sub.Buf => Sonic => 95.degree ./15' .sup. 617: 21/21 71.4% (1994) __________________________________________________________________________ For each study in Table 1 the processing protocol is abbreviated (see Footnote A), the number of clinical specimens evaluated is stated with th corresponding number of culture and amplification positive samples (see Footnote B), and then the "uncorrected" correlation to culture shown (see Footnote C). These 35 were chosen because a clear comparison between the amplification results and the culture results #was presented. The studie of Shankar, P. et al., Lancet 337:5-7 (1991); deWit, D. et al., Tubercle and Lung Dis. 73:262-267 (1992); and Kaneko, K. et al., Neurology 4:1617-1618 (1990) were omitted because no clear comparison on this basis could be made.
D. The Studies of Table 1
Examination of the data reported in the publications of Table 1 suggests that, regardless of system design or sample processing technique, there was a wide variation in results. These studies include literally all possible sources of specimen; including sputum, bronchial washes, pleural fluid, gastric aspirates, cerebrospinal fluid (CSF), urine, tissue biopsy, bone marrow, abscess and exudates, blood, serum, peritoneal fluid and feces. Two different amplification schemes were used: 30 studies used PCR exclusively, three studies used the isothermal, retroviral type, proprietary amplification scheme being commercialized by Gen-Probe (Jonas, V. et al., J. Clin. Micro. 31:2410-2416 (1993); Pfyffer et al., J. Clin. Micro. 32:918-923 (1994); Bodmer et al., J. Clin. Micro. 32:1483-1487 (1994)), and two studies compared the two amplification technologies (Abe, C. et al., J. Clin. Micro. 31:3270-3274 (1993); Miller et al., J. Clin. Micro. 32:393-397 (1994)). Thirty-three studies focussed on detection of MTB, one study focussed on M. avium diagnosis (Irula, J. V. et al., J. Clin. Micro. 31:1811-1814 (1993)), and one study pursued M. paratuberculosis detection (van der Giessen, J. W. B. et al., J. Clin. Microbiol. 30:1216-1219 (1992)). False negative results could be found in most specimen categories, regardless of target, processing technique or amplification technology.
The studies reported in Table 1 range in sample size from 7 to 1,166 specimens. Correlations with culture ranged from 3% to 100%. Nine of the 35 studies (26%) claim correlations of 100%. However, the majority, 7 of 9 (78%), involve sample sizes of less than 100 (n<100). Only two studies (Manjunath, N. et al., Tubercle 72:21-27 (1991); Pao, C. C. et al., J. Clin. Micro. 28:1877-1880 (1990)) used more than 100 specimens (however, Pao, C. C. et al., J. Clin. Micro. 28:1877-1880 (1990) may have reamplified in an effort to confirm negative samples: see footnote F in Table 1). Alternatively, 26 of the 35 studies (74%) show correlations of less than 100%. In this group, 20 of the 26 (77%) utilize sample sizes greater than 100 (n>100). It appears that there is an inverse relationship between the amplification-culture correlation and sample size: in general, the more samples included in a study, the lower the correlation.
In Table 1, 32 of the 35 studies (91%) show that amplification was able to detect the presence of Mycobacterial DNA in culture negative specimens (two of the three remaining studies used known positive specimens only). Two of these 32 more than double the number of culture positives (Irula, J. V. et al., J. Clin. Micro. 31:1811-1814 (1993); Pao, C. C. et al., J. Clin. Micro. 28:1877-1880 (1990)), and three very nearly double this number (Kolk, A. H. J. et al., J. Clin. Micro. 30:2567-2575 (1992); Manjunath, N. et al., Tubercle 72:21-27 (1991); Miyazaki, Y. et al., J. Clin. Micro. 31:2228-2232 (1993)). Thirty-one authors state directly, or reference the fact, that under ideal in vitro conditions their systems have the ability to detect the presence of 10 copies or less. The sensitivity of the remaining 4 range from 15 to 40 copies (Altamirano, M. et al., J. Clin. Micro. 30:2173-2176 (1992); Hermans, P. W. M. et al., J. Clin. Micro. 28:1204-1213 (1990); Pao, C. C. et al., J. Clin. Micro. 28:1877-1880 (1990); Soini, H. et al., J. Clin. Micro. 30:2025-2028 (1992)). Culture negative samples, that are positive by amplification, are more easily explained than culture positive-amplification negative samples: contrary to culture, PCR does not require viable organisms. For example, processing is known to kill the organisms, drug therapy may have already compromised viability, or low copy number combined with reduced viability might all contribute to the former class of samples. Regardless of system parameters, amplification should have superior sensitivity relative to either culture or smear.
Among the 35 studies, 8 different methods are used to process the raw specimens. Seventeen studies processed samples for both culture and amplification by treatment with N-acetyl-L-cysteine/NaOH (NALC), 6 studies used sodium dodecyl sulfate (SDS), 3 studies used sodium hydroxide (NaOH), 2 studies used dithiothreitol (DTT); and oxalic acid (OxAc), polyethylene glycol (PEG), and the ficoll-hypaque (Fic/Hyp) gradient method were each used once. One study actually compared processing by NALC with that of SDS (Pfyffer et al., J. Clin. Micro. 32:918-923 (1994)). Three studies avoided using the culture sediment and directly processed samples for amplification. Of the nine studies that obtained correlations of 100%, three used NALC, three processed specimens for amplification directly, one study used SDS, one used OxAc, and one used PEG. Keeping in mind that 7 of these studies utilized sample sizes of less than 100 (n<100), no conclusions can be drawn between how the raw specimens were processed, and perfect correlation. The study of Pfyffer et al., J. Clin. Micro. 32:918-923 (1994) compared NALC and SDS processing and concluded that neither method was superior.
Preparation of the processed sediment (or samples directly) for amplification falls into seven basic categories: (i) sixteen examples use variations on organic extraction and alcohol precipitation methodologies (org/ppt) as described by Maniatis, T. et al. ("Molecular Cloning A Laboratory Manual," Cold Spring Harbor Laboratory, New York (1982), pp. 458-463); (ii) two examples use an enzymatic lysis and boiling protocol (Lz/Prk.fwdarw.95.degree./15') as described by Higuchi, R. Amplifications 2:1-3 (1989), and six examples simply boil the specimen; (iii) ten examples use sonication (sonic); (iv) three use chaotropic agents and glass beads (GuSCN/Si) as described by Boom, R. et al., J. Clin. Microbiol. 28:495-503 (1990), and two use a similar protocol of binding the DNA to silica; (v) one example used sucrose gradient fractionation (Victor, T. et al., J. Clin. Micro. 30:1514-1517 (1992)); (vi) one example used Chelex-100 as described by de Lamballerie, X. et al., Res. Microbiol. 143:785-790 (1992); and (vii) one example (van der Giessen, J. W. B. et al., J. Clin. Microbiol. 30:1216-1219 (1992)) used the column chromatography procedure suggested by the manufacturer (Vary, P. H. et al., J. Clin. Microbiol. 28:933-937 (1990)). The sum is greater than 35 because the studies of van der Giessen, J. W. B. et al., J. Clin. Microbiol. 30:1216-1219 (1992), Victor, T. et al., J. Clin. Micro. 30:1514-1517 (1992), Wilson, S. M. et al., J. Clin. Micro. 31:776-782 (1993) and Abe, C. et al., J. Clin. Micro. 31:3270-3274 (1993), Miller et al., J. Clin. Micro. 32:393-397 (1994) and Pfyffer et al., J. Clin. Micro. 32:918-923 (1994) process all specimens by more than one protocol and present analyses for each method. Eight of the nine studies that claim 100% correlation isolate the DNA by organic extraction/alcohol precipitation methodologies. However, eight studies that also use organic extraction methodologies had correlations ranging from 55.9% to 98.4%. One study that claimed 100% correlation was processed by boiling (Kocagoz, T. et al., J. Clin. Micro. 31:1435-1438 (1993)). Alternatively, six studies using this same protocol reported correlations between 78.9% and 95.9%. Seven studies actually compare methods to prepare the culture sediment for amplification. Their conclusions differ as follows: Pierre, C. et al., J. Clin. Micro. 29:712-717 (1991)) selected organic extraction/alcohol precipitation; Wilson, S. M. et al., J. Clin. Micro. 31:776-782 (1993)) simply treated with chloroform; Folgueira, L. et al., J. Clin. Micro. 31:1019-1021 (1993) preferred the enzymatic lysis/boiling method; Kocagoz, T. et al., J. Clin. Micro. 31:1435-1438 (1993) and Sritharan, V. et al., Mol. Cell. Probes 5:385-395 (1991) chose the simple boiling method; and Forbes, B. A. et al., J. Clin. Micro. 31:1688-1694 (1993) and Buck, G. E. et al., J. Clin. Micro. 30:1331-1334 (1992) identified sonication as the optimal method (only the study of Kocagoz, T. et al., J. Clin. Micro. 31:1435-1438 (1993) achieved 100% correlation). It would appear that the occurrence of false negatives is not only independent of the protocol employed to prepare the sample for amplification, but there is controversy surrounding this issue as well.
Two studies actually compare amplification methods: Abe, C. et al., J. Clin. Micro. 31:3270-3274 (1993) show that the Gen-Probe method was marginally better than PCR (see footnote .kappa. in Table 1), while Miller et al., J. Clin. Micro. 32:393-397 (1994) determine the opposite (see footnote .mu. in Table 1). Apparently, neither amplification technique confers a significant advantage for clinical diagnosis of TB infections.
E. PCR Inhibitors
Of the papers reporting correlations less than 100%, 17 studies refer to amplification "inhibitors" as a contributing factor to false negatives (see those authors with a superscript .dagger. in Table 1). Blood (Panaccio, M. et al., Nucl. Acids Res. 19:1151 (1991)), feces (Allard, A. et al., J. Clin. Microbiol. 28:2659-2667 (1990)), sputum (Hermans, P. W. M. et al., J. Clin. Micro. 28:1204-1213 (1990); Shawar, R. M. et al., J. Clin. Micro. 31:61-65 (1993)) and urine (Khan, G. et al., J. Clin. Pathol. 44:360-365 (1991)) all contain PCR inhibitors. In addition, with respect to sputum, bronchial washes and tracheal aspirates, there is a direct correlation between the viscosity of the specimen (mucous content) and disease state: patients with advanced stages of tuberculosis have the most viscous sputum and these specimens have the highest probability of retaining amplification inhibitors. Hermans, P. W. M. et al., J. Clin. Micro. 28:1204-1213 (1990)) and Shawar, R. M. et al., J. Clin. Micro. 31:61-65 (1993)) show reductions in sensitivity of 5-20 fold, and 5 fold, respectively, in the presence of sputum.
Of the 42 methodologies presented for processing in Table 1 only twelve do not incorporate some form of buffer exchange. For example, organic extraction/precipitation, washing of the pellet, or protocols using chaotropic agents (GuSCN/Si), all require a buffer exchange at some point. Sonication of the sediment, however, does not require a buffer exchange. None of these twelve studies achieves 100% correlation, and nine within this group refer to inhibitors as a contributing factor to false negatives. Inhibitors appear to be derived from both the specimen and solutions used for processing, and both sources pose significant challenges to clinical implementation of amplification technologies.
F. Low Copy Numbers, Statistical Dropouts and "Unexplained" Results
Statistical dropouts, also referred to as "sample bias," are due to low copy number; in a sample with extremely low copy numbers, from which aliquots must be taken, there exists the possibility that some aliquots will contain no target. For example, if there are ten copies of the target in a milliliter, and ten 100 .mu.l aliquots are taken, target will be absent from some fractions. These aliquots, while being interpreted as false negatives, are "true amplification negatives." Eight studies in Table 1 describe a phenomenon that could be explained by this type of sample bias (see those authors' names with a superscript .sctn. in Table 1). As discussed below, this phenomenon is greatly exacerbated by aggregation.
Of the papers reporting correlations less than 100%, 15 refer to "low copy number" directly as a contributing factor to false negative results (see those authors with a superscript .dagger-dbl. in Table 1). However, 6 of these 15, plus 3 others, present examples where negative amplification specimens were both culture positive and smear positive (see those authors' names with a superscript .paragraph. in Table 1). The limit of detection of acid fast staining has been reported as 7,800 to 9,500 organisms per milliliter of sputum (Hobby, G. L. et al., Antimicrob. Ag. Chemother. 4:94-104 (1973); Yeager, H. et al., Amer. Rev. Resp. Dis. 95:998-1004 (1967)). Clarridge, J. E. et al., J. Clin. Micro. 31:2049-2056 (1993) present an extensive analysis of false negatives (see Table 7 of this reference). Of 37 false negative specimens analyzed in detail, 26 showed dropouts, while 11 were "true" PCR negatives. Nine of these 37 were smear positive: 4 of these 9 contained inhibitors, 3 were not tested for inhibitors, and 2 were found to be free of inhibitors. Of these last two, one was a true PCR negative. Shawar, R. M. et al., J. Clin. Micro. 31:61-65 (1993) also reported culture positive-smear positive-PCR negative specimens that were seen to be free of inhibitors. If the sample does not contain inhibitors, and is smear/culture positive, "low copy number" cannot be a realistic possibility. Shawar, R. M. et al., J. Clin. Micro. 31:61-65 (1993) refer to this class of false negatives as "unexplained."
G. Partitioning of Mycobacteria During Centrifugation
The buoyant nature of Mycobacterium was evident as early as 1924 (Andrus, P. M. et al., Am. Rev. Tuberc. 9:99 (1924)). Since then, several studies have highlighted the difficulty of sedimenting Mycobacteria (Hanks, J. H. et al., J. Lab. Clin. Med. 23:736-746 (1938); Hata, Jr., D. et al., Dis. Chest 18:352-362 (1950); Klein, G. C. et al., Am. J. Clin. Pathol. 22:581-585 (1952); Ratman, S. et al., J. Clin. Microbiol. 23:582-585 (1986); Rickman, T. W. et al., J. Clin. Microbiol. 11:618-620 (1980); and Robinson, L. et al., J. Lab. Clin. Med. 27:84-91 (1941)), and, in several instances, culturing the supernatant is advocated as standard practice.
While several studies report that the supernatant fractions contained smear positive material (Hanks, J. H. et al., J. Lab. Clin. Med. 23:736-746 (1938); Rickman, T. W. et al., J. Clin. Microbiol. 11:618-620 (1980)), another study showed that in 88.8% and 82.4% of all specimens centrifuged at 2,000 rpm and 3,000 rpm, respectively, the supernatant was culture positive (Klein, G. C. et al., Am. J. Clin. Pathol. 22:581-585 (1952)). In fact, this same study showed that in 2.2% and 2.7% of all specimens centrifuged at 2,000 rpm and 3,000 rpm, respectively, the sediment was culture negative while the supernatant was culture positive. Analyzing the supernatant fraction is still discussed in contemporary laboratory manuals (Kent, P. T. et al., "Public Health Mycobacteriology" in A Guide for the Level III Laboratory, U.S. Department of Health and Human Service, Centers for Disease Control, (1985) pp. 31-46; Sommers, H. M. et al., "Mycobacterium," in: Manual of Clinical Microbiology, E. H. Lennette et al., eds., 4th ed., Am. Soc. Microbiol., Washington, D.C. (1985), pp. 216-248).
The inverse relationship between sample size and correlation could potentially be explained by the buoyancy phenomenon. Larger sample sizes require batch processing. During batch processing the time it takes between work-up of the first and last specimens increases. As this time increases, buoyancy has a greater amount of time to exert an effect. It has been suggested that the source of this buoyancy is the high lipid content of these organisms (Silverstolpe, L. Nord. Med. 40/48:2220-2222 (1948)).
H. Influence of the Cell Wall and Surface Tension on Recovery of Mycobacteria
The nature of the cell wall of the Mycobacteria is responsible for their survival tenacity. Micrographs reveal a very complex structure 30-40 nm thick (Rastogi, N. et al., Antimicrob. Agents Chemother. 20:666-677 (1981)). As much as 60% of the dry weight of the cell wall is lipid (Joklik, W. K. et al., Zinsser Microbiology 20th edition, Appleton & Lange, Norwalk, Conn. (1992), pp. 499).
The cell wall of the Mycobacterium has three distinct layers: (i) the peptidoglycan, (ii) the arabinogalactan, and (iii) glycolipids (for a comprehensive review of cell wall structure see McNeil, M. R. et al., Res. Microbiol. 142:451-463 (1991)). Mycolic acids, which are extremely hydrophobic and consist primarily of hydrocarbon chains (.SIGMA.=C.sub.76 -C.sub.80), are used extensively in the construction of both the arabinogalactan and the glycolipid layers. The structure and species distribution of mycolic acids is reviewed in Takayama, K. et al., "Structure and Synthesis of Lipids," in: The Mycobacteria: a Source Book, Part A, G. P. Kubica et al., eds., Marcel Dekker, Inc., New York, N.Y. (1984), pp. 315-344. The Mycobacteria are essentially encased in wax.
M. tuberculosis form "cords" during growth (cords are clumps or aggregates of large numbers of organisms), and there is a direct relationship between MTB virulence and cord formation (Joklik, W. K. et al., Zinsser Microbiology 20th ed., Appleton & Lange, Norwalk, Conn. (1992), pp. 503). Organisms of the MAC complex have additional glycolipid (C-mycoside) components in their cell wall (the differences among the serovars is summarized in Brennan, P. J. Rev. Infect. Dis. 11(Supp. 2):s420-s430 (1989)).
In culture, while MTB complex organisms are seen to form dense clumps, M. avium grows in a more diffuse, single cell fashion (Dubos, R. J. et al., J. Exp. Med. 83:409-423 (1946)).
Dubos, R. J. Proc. Soc. Exp. Biol. Med. 58:361-362 (1945) observed that the hallmark pellicle growth of cultured MTB could be modified by the addition of the polyoxyalkylene derivative of sorbitan monostearate (Tween 60: CAS.RTM.No. 9005-67-8). This observation was later extended to show that other similar derivatives could cause MTB to exhibit "rapid," "diffuse" and "submerged" growth characteristics (Dubos, R. J. et al., J. Exp. Med. 83:409-423 (1946)). Tween 80 (CAS.RTM.No. 9005-65-6) was found to be the most active in that regard. These authors concluded that submerged growth was due to "wetting" of the cell surface. The term "wetting" is used exclusively in the context of surface tension. The implication of these studies was that pellicle growth resulted from surface tension between the waxy coat and the aqueous media, and that the addition of Tween 80 alleviated this physical interaction.
If surface tension confined the organisms to the surface, this, in combination with cording, could explain aberrant results: very sick patients are infected by organisms that have a propensity for cord formation. In addition, large cords produce smear positive results and cultures that turn positive fairly quickly. Large cords would also exacerbate sample bias because a large cord would distribute as a single copy, but have the potency of thousands of copies. Consequently, as suggested by Klein, G. C. et al., Am. J. Clin. Pathol. 22:581-585 (1952), the microorganisms would easily be poured off with supernatant fraction, thereby facilitating the sample bias phenomenon. In addition, cording would cause the bacteria to partition such that smear/culture positive and amplification negative results would occur.
If surface tension and aggregation were responsible for the anomalous results, and surface tension and aggregation could be overcome by the addition of nonionic detergents, then it seems logical that these reagents should improve correlation to culture: five studies in Table 1 use nonionic detergents to wash sediments prior to amplification (Clarridge, J. E. et al., J. Clin. Micro. 31:2049-2056 (1993); Irula, J. V. et al., J. Clin. Micro. 31:1811-1814 (1993); Kolk, A. H. J. et al., J. Clin. Micro. 30:2567-2575 (1992); Shawar, R. M. et al., J. Clin. Micro. 31:61-65 (1993); and Sritharan, V. et al., Mol. Cell. Probes 5:385-395 (1991)). The correlations to culture range from 78.9% to 95.5% in this subset. As early as 1941 it was recognized that agents that alleviated surface tension were impotent in enhancing recovery by centrifugation (Robinson, L. et al., J. Lab. Clin. Med. 27:84-91 (1941)). Therefore, the art teaches there clearly is no additional advantage to inclusion of these detergents in the wash buffer.
I. Problems Unique to MAC Complex Organisms
Of the 35 studies in Table 1, 27 use MTB specific sequences for amplification and/or detection. Six studies use genus specific primers, but the designs preferentially favor amplification of TB complex organisms. Only the study of Irula, J. V. et al., J. Clin. Micro. 31:1811-1814 (1993) focussed on M. avium, and only the study of van der Giessen, J. W. B. et al., J. Clin. Microbiol. 30:1216-1219 (1992) focussed on M. paratuberculosis. Irula used a different isolation technique, making comparisons difficult. However, while PBMC were isolated, they were in fact subjected to a wash step in Tris/EDTA/Triton X-100: if the PBMC had lysed during washing, the bacteria may have been discarded with the supernatant. Regardless, it might be expected that PBMC isolation would be an extremely effective means of capturing the organisms. The study of van der Giessen compared three PCR based systems (McFadden, J. J. et al., Mol. Microbiol. 1:283-291 (1987); van der Giessen, J. W. B. et al., J. Med. Microbiol. 36:255-263 (1992); Vary, P. H. et al., J. Clin. Microbiol. 28:933-937 (1990)) designed to detect M. paratuberculosis in bovine feces (one of these is a commercially available kit from IDEXX (Vary, P. H. et al., J. Clin. Micobiol. 28:933-937 (1990))). Their results were far worse than anything else presented in Table 1 and appear to be artificially low. These results indicate that MAC complex organisms, with their additional lipophilic components, present a further undefined complication to processing for amplification.
J. The Innate Character of the Mycobacteria has Thwarted Exploitation of Amplification Technologies
Clearly, there are two primary sources of false negative results. First, inhibitors are abundant in a variety of specimen types and the preparatory solutions also play a role in modifying the efficiency of the amplification reaction. The second category is due to the innate character of the Mycobacteria. While the source of these characteristics appears obvious, the influence of these idiosyncrasies on sensitivity is so prevalent that Noordhoek et al., J. Clin. Micro. 32:277-284 (1994) conclude: " . . . we will not speculate on the possible factors that might explain the extreme differences in sensitivity of PCR among the seven laboratories . . . . "
The cording, buoyant nature of these organisms causes them to partition in an inefficacious manner, and be poured off with the supernatant. An extreme example of this situation causes "unexplained" results: a sample that is culture positive and smear positive, but appears to be a true negative in the face of multiple amplifications, and does not contain inhibitors. Clearly, the source and nature of these characteristics remains to be fully explained. However, it is these phenomena, the basis of which resides in the innate qualities of these organisms, that the methods described herein solve.


An apparatus for dissociation of gas molecules which absorb light comprising a resonance optical cavity, a nozzle for introducing gas molecules into the resonance optical cavity in contact with the high velocity gas molecules, a focusing lens for focusing the laser beam at the point of contact of the laser beam and the high velocity gas molecules, a 100 percent reflecting mirror for receiving the laser beam after it passes through such molecules, a partially reflecting mirror positioned between the molecules for reflecting a portion of laser beam back into the laser and the remainder of the beam through the gas molecules to the 100 percent reflecting mirror.

In the conventional method of producing tetrafluoroethylene (TFE) chlorodifluoromethane is subjected to elevated temperatures in the order of 700.degree. to 800.degree. C. At this temperature, the reactants thermally dissociate into TFE and other products. It is necessary to cool the reaction products to subzero temperatures in order to effect separation and recovery of the TFE. The energy cost of the commercial process is very high because of the large amounts of heat required and the necessity of reducing the reaction products to very low temperatures.
Multi photon dissociation (MPD) of molecules which absorb light through the use of lasers has been used by many investigators, primarily for isotope enrichment. MPD has also been used to dissociate reactants into various products, however, this method has not found commercial acceptance because of the low efficiency of the dissociation process.
Trifluoromethane (CHF.sub.3) dissociates according to the following reaction steps: ##STR1##
The overall efficiency of the conversion of CHF.sub.3 with MPD to TFE is on the order of 0.03 to 0.12 percent. This efficiency is determined by multiplying together the three separate efficiencies which make up the MPD process; viz. pulsed laser wall plug efficiency, 3.0 percent; fractional absorbance of light by the reactant, 10-20 percent; and photon efficiency less than 100% , the energy required to break the chemical bonds of TFE produced divided by the total energy absorbed.
The low overall efficiencies of the MPD process have prevented the use of MPD for large scale commercial production of products such as TFE.
Use of the MPD process for the conversion of CHF.sub.3 to TFE would provide substantial energy advantage over the conventional chemical process. The laser method does not require heat input because dissociation occurs by selective absorption of laser light rather than by heating. Cooling to separate products in the MPD process is required, but, the energy consumption is less than in the conventional chemical process because the products are obtained at a much lower temperature.
It is desirable to provide an MPD process and apparatus in which the efficiency of the dissociation of molecules to form desired products is substantially increased.

Apparatus for the laser dissociation of molecules


A door positioner for holding open a side door of a vehicle body, in particular during a surface treatment of the vehicle body, includes a U-shaped bracket with a first limb, a second limb and a connecting element connecting the two limbs. The two limbs are deflectable elastically against one another in a shear plane. Arranged at the free end of the first limb is a fastening piece for the detachable fastening of the door positioner on an inner surface of the side door. Provided at the free end of the second limb is an actuating part for deflecting the second limb relative to the first limb. Located on the second limb between the connecting element and the actuating part is a fixing means for spring locking of the door positioner on a door spar of the vehicle body.

1. Field of the Invention
The invention relates to a door positioner for holding open a side door of a vehicle body, in particular during a surface treatment of the vehicle body.
2. Description of Related Art
In the production of vehicles, a body shell is first produced from individual parts. This is then treated in a surface treatment plant in different ways, e.g. by applying paints and other coatings. Generally the body shell is immersed in this process in immersion baths and rotated and/or moved in a translatory manner therein.
During the immersion process the side doors of the body shell should be slightly opened, so that all parts of the side doors and the door surrounds can be treated uniformly. An open position of the side doors is also favorable so that paints and other liquids can drain from the body shell more easily.
If a body shell with opened side doors is moved in an immersion bath, the dynamic pressures caused by the movement could lead to the side doors closing again or opening too far. To prevent this, door positioners are used, which hold the slightly opened side doors in a defined open position in the immersion baths and frequently also during processing in other processing stations.
Known door positioners basically comprise an L-shaped angle with a short limb and a long limb. The free end of the short limb bears a fastening piece, via which the door positioner can be screwed or otherwise fastened detachably to the inside of the side door. At the other end the short limb is connected via a leaf spring to the long limb, which in the fitted state extends into the interior of the body shell.
Arranged laterally on the long limb is a retaining notch, which in the open position of the side door engages in a rebate of the vertical door spar. Arranged at the free end of the long limb is an actuating part, with which the long limb can be swung out against the resistance of the leaf spring in a horizontal plane a few degrees from its resting position. Due to horizontal swinging of the long limb the retaining notch can be brought into engagement with the rebate of the door spar, in order to fix the side door in the open position. By renewed actuation of the actuating part this fixing can be released again, so that the side door can be closed or completely opened.
On the known door positioner the actuating part is situated relatively far in the interior of the body shell. If a robot is to be used for opening and closing the side door, this must reach with one robot arm relatively far into the interior of the body shell to be able to operate the actuating part. This is disadvantageous insofar as paint traces and other particles, which have settled on the robot arm, can detach themselves from this and fall down onto the door sill when the robot arm reaches into the body shell. Since the door sill is not generally clad on motor vehicles, such contaminations can result in expensive extra work on the body shell to guarantee an aesthetically perfect appearance.


This invention relates to compositions comprising anti-CD83 agonist antibodies and methods of treating autoimmune disorders (such as inflammatory bowel disease) with anti-CD83 agonist antibodies, as well as articles of manufacture comprising anti-CD83 agonist antibodies.

Inflammatory bowel diseases (IBD), such as ulcerative colitis and Crohn's disease, are characterized by chronic and relapsing inflammation of the intestine. Recent evidence indicates that IBD is due to a loss of tolerance to commensal intestinal flora in which the mucosal immune system can not differentiate between pathogens and commensal organisms. Therefore IBD patients develop immune recognition to normal flora which results in a gastrointestinal inflammatory response. The extent to which seroreactivity to normal flora develops is roughly correlated with disease duration and disease severity (Lodes et al., J Clin Invest., 113:1296-1306, 2004). More than 25 alleles have been identified that overlap between ulcerative colitis and Crohn's disease genetics (Umeno et al., Inflamm Bowel Dis. In press, 2011). These IBD genetic alleles converge on several common pathways that result in 1) Defects in epithelial integrity, 2) Defects in myeloid microbial responses and production of proinflammatory cytokines, and 3) Increased T-helper 17 cell and T-helper 1 cell responses. Studies show that most IBD patients have a spontaneous relapse and remission cycle where they achieve clinical remission at reasonable frequency but do not sustain remission for extended periods. This observation indicates the existence of as yet undefined mucosal homeostatic mechanisms that may drive the IBD remission phase (Schirbel et al., Expert Rev Gastroenterol Hepatol., 5(1):33-41, 2011). The identification of molecules and mechanisms involved in maintaining mucosal homeostasis would therefore be beneficial for driving as well as maintaining IBD remission.
CD83 is a highly conserved 45 kilodalton transmembrane glycoprotein that is predominantly expressed on the surface of dendritic cells and thymic epithelial cells. Additionally, CD83 is transiently expressed on the surface of other activated cells of the immune system and can also be found in a soluble form. Structural analysis of the predicted amino acid sequence of CD83 demonstrates that it is a member of the immunoglobulin super family indicating a role in the immune system. Literature reports suggests that soluble CD83 (sCD83) may play an immunomodulatory role due to the observation that sCD83 released by HCMV-infected mature dendritic cells inhibits T cell proliferation (Senechal et al., Blood., 103(11):4207-4215, 2004) and that sCD83 has a morphologic effect on the dendritic cell cytoskeleton (Kotzor et al., Immunobiology., 209(1-2):129-140, 2004). However, no ligand has been identified for CD83 and the function of CD83 remains poorly understood. Interestingly, CD83 gene expression is downregulated at the mucosal surface in human Crohn's disease (Silva et al., Dig. Dis. Sci., 53(7):1917-1928, 2008), suggesting that CD83 may be involved in the maintenance of mucosal homeostasis and therefore be a therapeutic target for modulating the immune system in IBD.
All references cited herein, including patent applications and publications, are hereby incorporated by reference in their entirety.

Use of anti-CD83 agonist antibodies for treating autoimmune diseases


Various antenna designs that are small and cost effective are disclosed. In one design, the antenna is a bent and folded monopole antenna. In another design, the antenna is a folded and tapped monopole antenna. In yet another design, the antenna is a folded, bent, and tapped monopole antenna. The antennas may be part of a system using two back-to-back symmetric antennas. The antennas may be part of a modem, such as a wireless computer modem or a wireless handset.

The increasing use of wireless communications leads to a need for clearer transmissions while reducing the overall size of communications devices. The move to smaller communications devices has caused a need for new antenna designs and systems to use them. Traditional ceramic chip antennas are small but costly. It is thus desirable to implement a small, cost-effective antenna.

Folded monoplole antenna, bent, tapped, or both, and systems incorporating same


Disclosed is a beverage container including a container body (1) and a floating object (2) for reducing the sloshing amplitude of the beverage, the floating object (2) is set in the container body (1) and floats on the surface of the beverage in the container body (1), the floating object (2) is larger than an opening (3) of a top cover (4) of the container body (1), the floating object (2) is provided with at least one through hole (21) for preventing the floating object (2) from blocking the opening (3) of the top cover (4) of the container body (1).

In recent years, with the improvement of the quality of people's life and the acceleration of the pace of life, the dietary structure of consumers and their eating habits have changed. People start to pursue simple and fast diet, which has also become a new consuming trend. Due to the awareness of living a healthy life, the market starts to widely sell milk, juice drinks, carbonated beverages and tea beverages. And young people are the main consumers. The containers of these beverages are different. Among them, cans and special paper boxes are most widely used. Cans and the boxes are respectively provided with a dispense area for sealing the beverage container and a pull tab connected to the dispense area. The beverage container is opened through pulling the pull tab to separate the dispense area from the container.
Usually, while pulling the pull tab or walking with the beverage container after it is opened, the container shakes. When the beverage in the container oscillates with the shaking amplitude of the container, a resonance effect occurs, and the fluctuation range and impact force of the beverage in the container are further increased. As a result, the beverage in the container is easily spilled, which causes inconvenience to drinking.

Beverage container and method for preventing spillage of beverages


A portable garbage can comprising a cylindrical can body, an annular cap receiver combined with the can body, two half caps swingably combined with the cap receiver, and an inverted L-shaped handle combined with the cap receiver and having a bottom square engage portion to insert in an inner recess of a square plate fixed on one side of the cap receiver, two opposite saw teeth provided in the inner opening of the square engage portion to engage and rotate a stationary tooth respectively fixed on one side of the two half caps to swing open or close an upper opening of the cap receiver combined with the can body for throwing garbage therein.

A known garbage can shown in FIG. 1 comprises a can body 10, a stationary half cap 11 formed on the can body and a movable half cap 12 to close a half opening. When garbage is to be thrown in the can, the half cap is to be moved open with hand. Then the half cap is pulled to close up the opening with hand. But the opening is not so large enough to let garbage thrown, so garbage may often touch or bump against the half cap 11 or the upper curved edge of the can body 10. In addition, the half cap 12 may easily become smeared by hand for opening it, and the can has no means for carrying or lifting up, not convenient for moving it, and if worse, the can body may be fallen down to the ground by accident, with the garbage accumulated therein scattering around.

Jong-Juang Tzyy, Jea-Jong Tsuen, Shiah-Yng Shiang, Tainan Shien


A high pressure automatic lime mud precoat filter cake heel removal system provides for steady kiln operation. The system provides multiple points of washing located such that washing of the entire filter can be accomplished in a short period of time. Each washing point comprises preferably two nozzles at predetermined angles to optimize washing of the filter media. The system is reciprocatively driven by a fractional horsepower gear motor connected to a torque converting coupling which is, in turn, connected to a threaded jack shaft or lead screw. The gear motor is controlled by a dual speed reversing variable frequency drive; limit switches automatically control cycling of the system in conjunction with logic circuits and sensors. This system is variably controlled so that cycle times may range optionally from approximately one to approximately five hours in length.

1. Field of the Invention
This invention relates to the field of vacuum filters. More specifically, this invention relates to the field of drum lime mud filters and an apparatus to improve cleaning and efficiency thereof.
2. Prior Art
Lime mud filtering has been known for a substantial number of years and drum type filters have been favored for many years as well. Due to the extensive interest in this field, many modifications and other devices have been invented and employed with lime mud drum filters. One such arrangement has been proposed by Esko Mattelmaki in U.S. Pat. No. 5,149,448. '448 discloses a drum filter for thickening lime sludge in which the lime sludge is thickened on a filter layer having a precoat formed of the lime sludge itself. The device is also provided with high pressure liquid nozzles which continuously reciprocate in the longitudinal direction of the drum. The nozzles are located under the doctor blade on the descending side of the drum. The reference teaches to mount two high pressure nozzles in that location in such a way that one of the nozzles breaks the precoat so that it drops into the vat and that the other nozzle washes the filter cloth clean. While the device does clean the filter it suffers from the drawback of increased wear on the filter medium due to unnecessary overcleaning of the filter element. Moreover, the device of the prior art requires a long cleaning time and, therefore, accelerates motor and drive mechanism wear.
Another prior art device is disclosed in U.S. Pat. No. 5,470,472 to Baird et al. The disclosure teaches a non-traversing cleaning mechanism located on the descending side of the filter drum. Baird et al. teaches that the jets should be angled with respect to the filter media by selecting an angle of from 0 to 45 degrees and more preferably from 0 to 30 degrees measured from a vertical reference.


Adjustable stops are mounted on a rotating rod, and one pivot arm of a linked pair extends into an interference position such that one stop will strike it. The linkage draws the second arm into interference position if the first arm is displaced from it. Each pivot arm will brake the stop if struck from one direction and will be displaced, drawing in the other pivot arm, if struck from the opposite direction. The pivot arms react oppositely to being struck from each respective direction of rotation. Thus, continued rotation of the rod in one direction results in a stop striking a pivot arm suitably to brake the stop, with the permitted arc of rotation determined by relative adjustment of the stops.

1. Field of the Invention
The invention generally relates to brakes. More specifically, the invention relates to rod or wheel brake, especially to an intermittent brake. A positive lock brake is applied to a rotating rod, such as to an automotive steering wheel shaft. The mechanism permits the shaft to rotate through a limited, preselected arc. At the end of the preselected arc, the mechanism applies a positive lock in the direction of rotation. A mechanism of this type is useful in conducting dynamic testing of vehicle performance.
2. Description of Related Art Including Information Disclosed Under 37 CFR 1.97 and 1.98
Evaluating the performance characteristics of a car, truck, other land vehicle, or water vehicle can require performing repeated dynamic evaluations. Often such testing requires that an expert test driver perform a defined maneuver, and such a maneuver may be repeated a plurality of times under similar or controlled conditions. In order to aid the test driver in accurately repeating a test, it is desirable to limit certain aspects of vehicle performance. These limitations may include, for example, speed and available degree of steering wheel rotation. Every vehicle has inherent maximum limits for these factors; but for testing purposes, it is desirable to apply limits other than inherent maximums. As one example, for accident reconstruction it may be desirable to drive the vehicle under conditions similar to those present at the time of the accident.
One desirable limitation in such evaluations is the permitted arc of rotation for the steering wheel. Only in the unusual case is maximum available arc of steering rotation to be used in an evaluation. Typically, a test will require a preselected arc of steering wheel rotation less than the maximum. This arc often is less than two turns from neutral wheel position. Because the arc may be more than one full revolution, a simple abutment stop cannot be used in every situation.
Computerized steering systems can control and repeat any specified pattern of steering wheel rotation. Such systems cannot be used in every case. A computerized system often is designed for use with only a specific vehicle or type of vehicle. When evaluating a specific vehicle that has been involved in an accident, it may be impractical or impossible to find and install a computerized system suitable for use with the selected vehicle. Further, computerized steering systems tend to be technically complex and correspondingly costly, which serves as a practical prohibition against using them in many testing situations.
It would be desirable to have a widely adaptable steering limiter that can be simply and readily installed on a test vehicle. Further, it would be desirable to have a steering limiter of this type that is adjustable to permit a variably selectable angle of maximum steering wheel rotation.
Various devices appear in United States patents for limiting rotation of a wheel or shaft.
U.S. Pat. No. 4,751,986 to Takahashi shows a rotor that is provided with a stop. An arcuate stator is spaced from the rotor and carries two stops, spaced apart by a specified angle. A ball circulates in the gap between the rotor and stator and between the two stops of the stator. The rotor can rotate in either direction. The rotor stops when the stop on the rotor has pushed the ball against either of the stops on the stator. From one extreme to another, the rotor can rotate between one and two rotations. This system could offer an ability to control available rotation either by starting the ball in a predefined starting position between the stops or by adjusting a spacing between the two stops on the stator. However, this system is not readily adapted to be installed on any selected vehicle. Further, adjustment requires considerable pre-fabrication and complexity.
U.S. Pat. No. 4,191,301 to Hickman et al. shows a rotating main shaft that carries an extending lug. On each revolution, the lug strikes a boss on a neighboring counting wheel, advancing the wheel by a partial turn. One of the bosses on the counting wheel eventually will reach and contact a fixed stop lug that prevents further turning of the counting wheel. This stop event also prevents further rotation of the main shaft in the initial direction. The number of bosses on the counting wheel determines how many times the shaft can rotate in one direction before the stop event. As taught in the patent, this system offers no fine adjustment of less than one turn of the main shaft. A possible, but untaught, modification to increase fine adjustment is to provide multiple extending lugs on the shaft to reach the stop point after one or more partial turns. However, this type of adjustability requires specific pre-fabrication to conform to each different degree of desired steering wheel rotation. Thus, this system is not well suited for variable testing.
U.S. Pat. No. 2,746,573 to Hastings shows a two-sided system for allowing more than one turn between the two sides of the system. One side of the system is a spiral spring that provides a spiral groove pathway between the spring walls. The second side of the system provides a follower block that is slidable on a radial pathway. As the two sides relatively rotate, the follower block moves radially in the spring grooves until it reaches a stop at either end of the pathway. It is not evident how this system could be applied to a pre-installed steering shaft and variably adjusted for a selected degree of wheel angle.
U.S. Pat. No. 2,744,416 to Feigin shows a system of fixed stops on two meshing gears with different numbers of teeth. Due to differential gear ratios, after a determinable number of rotations, the fixed stop on each gear meets the fixed stop on the other. This technique can be used to predetermine almost any number of rotations. However, this system is not well suited for use on a pre-installed steering shaft. It would present problems in installing a gear over the shaft, and it would require suitable differential gears for each different selected steering angle. These limitations cause this system to be poorly suited for use on different vehicles and for variable steering angles. U.S. Pat. No. 5,239,490 to Masaki et al. shows a computerized electronic apparatus for precisely detecting the rotation of a steering wheel and steering shaft. The apparatus employs a microprocessor to analyze the detected signals and operate an electronic power steering system or other rotating device. This system is applied for the purpose of controlling rotation in response to inputs, rather than to limit rotation. Nevertheless, changing this mode of operation may be possible. This patent is an example of a computerized steering system, mentioned above. The chief important limitations are high cost, difficult or impossible adaptability to different vehicles, and lack of suitability for use when a test driver is controlling the vehicle.
U.S. Pat. No. 3,055,235 to Turley shows a planetary gear system that provides indexing motion through a specified arc. This patent shows general background. As evident from the scope of the prior art, suitable devices of the desired kind are unknown. While several mechanisms perform a related function, they are not sufficiently versatile in their applications and structures to be adapted for mounting on the steering system of substantially any test vehicle. As a result, the need to provide a positive and variable rotation lock for dynamically testing substantially any selected vehicle is not reasonably met.
To achieve the foregoing and other objects and in accordance with the purpose of the present invention, as embodied and broadly described herein, the method and apparatus of this invention may comprise the following.


An aluminum alloy material contains Si: 1.0 mass % to 5.0 mass % and Fe: 0.01 mass % to 2.0 mass % with balance being Al and inevitable impurities, wherein 250 pcs/mm2 or more to 7×105 pcs/mm2 or less of Si-based intermetallic compound particles having equivalent circle diameters of 0.5 to 5 μm are present in a cross-section of the aluminum alloy material, while 100 pcs/mm2 to 7×105 pcs/mm2 of Al-based intermetallic compound particles having equivalent circle diameters of 0.5 to 5 μm are present in a cross-section of the aluminum alloy material. An aluminum alloy structure is manufactured by bonding two or more members in vacuum or a non-oxidizing atmosphere at temperature at which a ratio of a mass of a liquid phase generated in the aluminum alloy material to a total mass of the aluminum alloy material is 5% or more and 35% or less.

When manufacturing a structure, such as a heat exchanger, which employs an aluminum alloy material as a constituent member, it is required to bond the aluminum alloy materials to each other or the aluminum alloy material to another different type of material. Various methods are known as bonding methods for the aluminum alloy material. Among the various methods, a brazing method (hard soldering method) is used in many cases. The brazing method is used in many cases in consideration of, e.g., an advantage that strong bonding can be obtained in a short time without melting a base material. As methods for manufacturing a heat exchanger, etc. by employing methods of bonding the aluminum alloy material with the brazing, there are known, for example, a method of using a brazing sheet on which a brazing filler metal made of an Al—Si alloy is clad, a method of using an extruded member coated with a powdery brazing filler metal, and a method of assembling various members and then additionally applying a brazing filler metal to portions that are to be bonded (Patent Documents 1 to 3). Moreover, "Chapter 3.2 Brazing Alloys and Brazing Sheets" in Non-Patent Document 1 explains the clad brazing sheet and the powdery brazing filler metal in detail.
Up to date, various brazing methods have been developed in the field of manufacturing of a structure using an aluminum alloy material. In relation to vehicular heat exchangers, for example, when a fin member is used in a single layer, there have been utilized a method of using a brazing sheet made of a tube material sheet on which a brazing filler metal is clad, and a method of additionally coating Si powder or a Si-containing brazing metal over the tube material sheet. On the other hand, when the tube member is used in a single layer, there has been utilized a method of using a brazing sheet made of a fin material sheet on which a brazing filler metal is clad.
Patent Document 4 discloses a method of employing a single-layer brazing sheet instead of the above-mentioned clad brazing sheet. That method proposes the use of the single-layer brazing sheet for a heat exchanger as a fin member and a tank member of the heat exchanger.

Aluminum alloy material, aluminum alloy structure, and manufacturing method for same


A method of verification, in a taximeter or a travel distance counter, of tariff defining points in time which are dependent upon a real time clock, includes accelerating to a maximum relevant chronological continuous switching of the taximeter or the travel distance counter by executing a maintenance program in such a way that a chronological work cycle or timing of an evaluating program is effected in a time compressed form for triggering of tariff defining functions. The apparatus includes means for effecting the method.

Known taximeter devices or travel distance counters have an internal clock with a calendar output, which serves to trigger functions predetermined by tariff or fare parameters. These devices or counters can be automatic tariff changeovers, for instance, for the day to a night tariff or other equivalent changeovers.
In order to verify switchover instants for parameter changes, it is necessary to manually set the clock and the calendar immediately before the expected changeover instant, and then to await the reaction or to institute active tests with the aim of finding, for instance, a tariff effect before and after the actual switchover instant.
A series of rtc--standard building blocks provides for the possibility of accelerating the normal sequence in the counters for the purpose of a check of the correct transfer between the second, minute, hour and calendar counters by bridging over the prescaler chains for downscaling the typical quartz oscillator frequency.
However, such an acceleration possibility is unavailable to the time defined taximeter functions.

Method and arrangement for verification of tariff defining points in time in a taximeter


In a vibration preventing damper forming method and a vibration preventing damper integrally formed with a mechanical chassis according to the present invention, an opening side end portion 9e of a damper housing 9 and a vibration preventing damper forming portion of a mechanical chassis 6 are made of a resin. The damper housing 9 is mounted on the vibration preventing damper forming portion by the fixture of the resin portions. Thus, a vibration preventing damper 8 is integrally formed with the mechanical chassis 6. For this reason, in supporting in a vibration proof manner and in a floating matter the mechanical chassis 6 within an interior of a casing 10 by means of the vibration preventing damper 8, it is possible to improve the working efficiency in mounting the mechanical chassis 6 to the casing 10 and to cope with the requirement of miniaturization of a mounting space between the interior of the casing 10 and the mechanical chassis 6.

1. Field of the Invention
The present invention relates to a vibration proof or preventing damper, and more particularly to a vibration preventing technology suitable for equipments such as automotive audio systems or portable personal computers provided with a reading mechanism for reading data from a recording medium such as a CD, a CD-ROM and a hard disc in a non-contact manner.
2. Description of the Related Art
In the equipment provided with such non-contact type reading mechanism for the recording medium in which the vibrations should be prevented, a variety of vibration proof means have been conventionally provided in order to attenuate vibrations that adversely affect the readout of the data. FIGS. 5 and 6 schematically show reproduction devices used in, for example, personal computers or automotive audio systems. These reproduction devices are adapted to optically read out music data from a music CD and reproduce them. A mechanical chassis 3 provided with a motor 2a, an optical pickup 2b, a disc table 2c and the like is installed in an interior of a casing 1. The mechanical chassis 3 is supported elastically within the interior of the casing 1 by vibration preventing dampers 4 and 5 in a vibration preventing manner so that the vibration hardly would be transferred.
Such vibration preventing dampers 4 and 5 are typically shown in two types as shown in FIGS. 5A to 5C and 6A to 6C. The device shown in FIGS. 5A to 5C is an insulator type damper in which the vibration preventing damper 4 is composed of an elastic cylindrical member 4a made of rubber or the like and each of the engagement recess grooves 4b provided circumferentially on a side surface is fitted inside of a C-shaped engagement projection 3a of the mechanical chassis 3. Then, when amounting screw N1 is inserted into the hollow interior of the elastic cylindrical member 4a and threadedly fixed to the casing 1, the mechanical chassis 3 are supported elastically at a plurality of positions by the vibration preventing dampers 4 within the casing 1 in a vibration preventing manner.
Further, the device shown in FIGS. 6A to 6C is of a type in which viscous fluid is sealed in the interior and the vibration is attenuated by utilizing the viscous fluid resistance of the viscous fluid. This vibration preventing damper 5 has a bottomed cylindrical agitating sleeve portion 5a and is provided with a damper housing 5e consisting of an elastic wall portion 5b to elastically deform in the three-dimensional directions to suppress the floating movement of the agitating sleeve portion 5a and to expand so as to surround the agitating sleeve portion 5a and a circumferential all portion 5d made of a hard resin, fixed at one end to an elastic wall portion 5b and having an outward flange 5c at the other end. A viscous fluid 5f is filled in the interior of the damper housing 5c and sealed by fixing a lid member 5q to the damper housing 5e to form the vibration preventing damper 5. Then, when the vibration preventing damper 5 is mounted on the mechanical chassis 3, a support rod 3b projecting from the mechanical chassis 3 is inserted into the agitating sleeve portion 5a and held thereat. Further, when the vibration preventing damper 5 is mounted on the casing 1, a mounting screw N2 is inserted into a screw hole formed in the lid member 5g and then screwed to the casing 1. Thus, the mechanical chassis 3 are elastically supported at a plurality of positions by the vibration preventing dampers 5 within the interior of the casing 1 in the vibration preventing manner.
As described above, the excellent vibration attenuating effect by the vibration preventing dampers 4 and 5 is exhibited, whereby the vibration would be hardly transferred from the casing 1 from the mechanical chassis 3 elastically supported within the interior of the casing 1. However, there are some problems. One of them is a degradation in working efficiency when the chassis is mounted on the casing 1.
Namely, in any conventional example shown in FIGS. 5A to 5C and 6A to 6C, the mechanical chassis 3 and the vibration preventing dampers 4 and 5 are formed into separate pieces. For this reason, in order to complete the mounting work between the mechanical chassis 3 and the casing 1, in the case of the conventional example shown in FIGS. 5A to 5C, in addition to the mounting work of the vibration preventing damper 4 to the mechanical chassis 3, a screwing step using a much larger number of mounting screws N1 is required. Accordingly, the number of steps is increased and the steps are troublesome. Further, in the case of the conventional case shown in FIGS. 6A to 6C, in addition to the mounting work between the vibration preventing damper 5 and the mechanical chassis 3, it is necessary to perform the screwing work between the vibration preventing damper 5 and the casing 1 using a plurality of mounting screws N2. Further, in this case, a large number of steps are required.
Further, in addition to such degradation in working efficiency, there is a problem in coping with a miniaturization for the mounting space. The tendency of miniaturization of the equipment, i.e., the demand for downsizing is increasing day by day. Such demand is remarkable in the field of automotive audio systems or notebook personal computers. Namely, in the case of the conventional examples shown in FIGS. 5A to 5C and 6A to 6C, if the casing 1 is miniaturized, correspondingly, the mechanical chassis 3 is also to be miniaturized. As a result, the mounting space for the vibration preventing dampers 4 and 5, that is, a gap between the inner wall of the casing 1 and the mechanical chassis 3 is also narrowed. Accordingly, in order to meet this miniaturization requirement, in the case of the vibration preventing damper 4 shown in FIGS. 5A to 5C, it is inevitable to reduce a height of the cylindrical elastic member 4a. If so, there are some cases where it is impossible to realize the vibration attenuation performance that is required. Further, in the case of the vibration preventing damper 5 shown in FIG. 6, if the circumferential wall portion 5d is more decreased and the agitating sleeve portion 5a is more shortened to thereby decrease the filling amount of the viscous fluid 5f for the miniaturization, it is possible to meet the requirement for the miniaturization. However, also, in this case, it is inevitable to sacrifice the vibration attenuation performance.

Vibration proof damper forming method and damper integration type mechanical chassis


A substrate (100) is a light-transmitting substrate. A light-transmitting first electrode (110) is formed over the substrate (100). An insulating layer (150) is formed over the substrate (100) and the first electrode (110) and includes an opening (152) overlapping the first electrode (110). An organic layer (120) is located within at least the opening (152). A light-transmitting second electrode (130) is formed over the organic layer (120). An intermediate layer (200) is formed in at least a portion of a region of a lateral side of the first electrode (110) overlapping the first electrode (110). A refractive index of the intermediate layer (200) is between a refractive index of the substrate (100) and a refractive index of the first electrode (110).

In recent years, there has been progress in the development of light-emitting devices using an organic EL element as a light-emitting unit. The organic EL element has a configuration interposing an organic layer between a first electrode and a second electrode. Such a light-emitting device may require transparency depending on its application. In order to make the light-emitting device transparent, the first electrode and the second electrode may be formed of a transparent conductive material.
On the other hand, in a case where the light-emitting device is made transparent, the edge of an electrode of an organic EL element may become conspicuous. In consideration thereof, Patent Document 1 discloses providing a correction layer in a region where no cathode is formed, the correction layer having the same material and thickness as those of the cathode. As an example of the cathode, a thin film of a metal such as magnesium, aluminum, or calcium having a low work function is disclosed. The correction layer is formed on the cathode with an insulating layer interposed therebetween.
Meanwhile, Patent Document 2 discloses a substrate provided with a transparent electrode, composed of a refractive index control layer and a transparent electrode layer formed in order on a transparent substrate. In Patent Document 2, the average refractive index of the refractive index control layer is 1.45 to 1.60. In addition, the refractive index control layer is formed as a laminate of two or more kinds of layers, the entire film thickness being 1,000 to 2,500 nm.

This patent is subject to a terminal disclaimer.

Light-emitting device having intermediate layer located over interconnect


The invention relates to a method for testing the plausibility of output signals (u1, u2) of a resolver (21), by means of which an angular position of two elements (3-7) of a machine (R) can be determined in relation to each other.

So called resolvers are used in order to determine the angular position of two links of a machine which can be rotated in relation to one another with respect to an axis of rotation. One example of a machine with a link that can be rotated in relation to itself is a robot. Robots in general are handling machines which are equipped for the automatic handling of objects with useful tools and are programmable in several axes of movement in particular with regard to orientation, position and work routine. Robots usually have a robotic arm with several links and programmable controllers (control devices) which control or regulate the movement sequences of the robotic arm during operation. The drives are e.g. electrical drives and the links are pivoted in relation to one another with respect to axes of rotation.
Resolvers are known to a person skilled in the art, e.g. from EP 2 211 148 A2. A resolver is defined as an electro-magnetic transmitter for conversion of the angular position of two components moved in relation to one another to electrical output signals. Two components moved in relation to one another are for example two links pivoted with respect to an axis of rotation or a rotor of a motor that rotates relative to the stator of the motor. The output signals are as a rule two sinusoidal electrical signals shifted by 90 degree phases, in particular two sinusoidal electrical voltages shifted by 90 degree phases, on the basis of which the angular position modulo n can be determined, wherein n is the so-called number of pole pairs of the resolver which arises from the arrangement of coil windings and is a small positive natural number, and can also be 1.

Method for testing the plausability of output signals of a resolver


The invention aims at providing a pair of binoculars making it possible to easily perform a focusing operation and a diopter adjustment, and efficiently perform an assembling operation. The present invention provides a pair of binoculars having a pair of supporters for supporting adjustment lenses provided between a pair of ocular optical systems and a pair of objective optical systems so as to be capable of moving forward and backward along the optical axis, a focusing gear for performing a focusing operation by moving said pair of supporters forward or backward at the same time in the direction of optical axis, a diopter adjuster for performing a diopter adjustment by moving one supporter forward or backward in the direction of optical axis as keeping the other supporter stationary, and a focusing and diopter adjusting unit having a changeover gear for changing over the focusing gear and the diopter adjuster to each other.

1. Field of the Invention
The present invention relates to a pair of binoculars, and more particularly to a pair of binoculars making it possible to easily perform a focusing operation and a diopter adjustment, and efficiently perform an assembling operation.
2. Prior Art
Up to now, there has been demanded a structure making it possible to easily perform a focusing operation and a diopter adjustment in an optical device such as a pair of binoculars and the like.
As a pair of binoculars making it possible to easily perform a focusing operation and a diopter adjustment, there has been proposed, for example, "a pair of binoculars in which two pipe members (1, 1′) of said pair of binoculars are connected with each other through two bridges (2, 3) each having a joint in order to adjust an interpupillary distance, said two bridges turn around the axes of joints (4) and are disposed distantly from each other by a specified distance with no connection by the central axis in order to form a penetrating portion (6), said two pipe members (1, 1′) have a focusing gear (8) being axially movable for focusing, said focusing gear is operated by a common rotary knob (12) and a gear, said gear extends through one of said bridges (2 or 3) having joints from said rotary knob (12) to said focusing gear (8), and an interpupillary adjustment has no influence on focusing" (see patent literature 1).
[Patent literature 1] Japanese Patent Laid-Open Publication No. 2000-56237 (claim 1)
Hereupon, in the invention described in said patent literature 1, there is performed a focusing operation as described as "A rotary knob is turned at a position where it has been inserted for focusing a pair of binoculars. The turning of the rotary knob is transferred from a rotary knob shaft directly to a friction disk and a projection. The projection engages the friction disk with a helical gear. The turning of the rotary knob is converted into an axial movement of a stop member by the helical gear, and an angular lever moves a focusing gear in the axial direction by an end portion having a fork. Since when a pipe member is turned around the axis the angular lever slides on a conical stop member at its end portion without the action of a gear for converting the turning of the knob into an axial movement of the focusing gear, in a pair of binoculars of the present invention a focusing operation is performed independently of an interpupillary adjustment." (See paragraphs [0030] and [0031] in patent literature 1.)
And in the invention described in patent literature 1 described above, a rotary knob is turned around a joint axis by moving radially an angular lever in order to perform a diopter adjustment. As a result, the turned rotary knob moves a fixed focusing gear in the axial direction.
As described above, in the invention described in patent literature 1, since the amount of movement of a focusing gear in the axial direction is adjusted in order to perform a diopter adjustment, it is necessary to perform such adjustment at plural points as positional adjustment between an angular lever and a supporter, positional adjustment between a coupler and a joint axis, and the like. In the invention described in patent literature 1, therefore, there is a problem that since the above-mentioned members also need to be adjusted in position when assembling the whole of a pair of binoculars it is not possible to efficiently perform the operation of assembling.


A CPU deals, as valid data, with demodulated data obtained by synthesizing demodulated data obtained by reading a first data block of an upper stage with demodulated data obtained by reading a second data block of the upper stage, and demodulated data obtained by synthesizing demodulated data obtained by reading a first data block of a lower stage with demodulated data obtained by reading a second data block of the lower stage, as far as both of the demodulated data obtained by reading the first data block of the upper stage of two column codes structured by arranging WPC codes up and down at two columns and the demodulated data obtained by reading the first data block of the lower stage, contain a portion corresponding to data characters of the second data block, and as far as these sets of demodulated data are different from each other.

1. Field of the Invention
The present invention relates to a bar code reader and a bar code reading method suitable for reading two column bar codes structured by arranging bar codes at two columns, and to a computer readable medium for storing a program for such a reading process.
2. Description of the Related Art
In recent years, it has been generalized that commercial goods are managed by bar codes as typified by a POS system in the sector of distribution industries etc. For example, in the POS system in a shop, pieces of information on classifications and sales prices etc of the goods are coded in a format of the bar code and printed on the goods. The bar code is read at the check-out counter, and a payment is made based on the thus read information. Then, the number of sold goods is counted in real time, and a result of count serves for an inventory management and a stocking management.
The above bar code is roughly classified into a WPC code such as a JAN code, a UPC code and an EAN code, and into a variable length second code. Among these codes, the WPC code has a start guard bar (SGB) added to a left end thereof, a center bar (CB) inserted in a middle portion thereof and an end guard bar (EGB) added to the right end thereof. A left (first) data block consisting of six or four data characters is provided between the start guard bar and the center bar, and a right (second) data block consisting of six or four data characters is provided between the center bar and the end guard bar.
Pattern of these guard bar and the center bar are predetermined by the specification, and therefore a bar code reader is capable of detecting these guard bars and center bar.
Further, the bar code reader is capable of reading the data character (a coded numerical values or symbolic minimum unit) of the bar code on the basis of the guard bars and the center bar. There are three reading methods, i.e., a continuous reading method, a block reading method and a division reading method. The continuous reading method is such that when detecting the two guard bars and the center bar by one bar code scanning process, the data characters interposed between the two guard bars are demodulated as a bar code. Further, the block reading method is that the block of the data characters is recognized as a bar code though the block is interposed by one guard and the center bar, and, after separately demodulating the two blocks, demodulated data of these respective blocks are synthesized (integrated), thereby reproducing the data modulated into the single bar code. Moreover, according to the division reading method, even a data character string contiguous to the single guard bar or the center bar at the minimum is recognized as a bar code, and fragments of the demodulated data of those data characters that have been separately read are synthesized, thereby reproducing the demodulated data corresponding to the whole single bar code. Namely, the block reading method and the division reading methods are implemented in such a case that the whole bar code ranging from the start guard bar to the end guard bar can not be read by one scanning process.
Incidentally, the WPC code has its own limit in terms of a quantity of expressible information because of being a fixed length code. Hence, for the purpose of increasing the quantity of expressible information while keeping the code system of the WPC code, there has hitherto been implemented a method in which the WPC codes are arranged at two columns (stages) and treated as one set of bar codes, and one piece of information is expressed by the bar codes arranged at the two columns (hereinafter referred to as a "two column bar code"). FIG. 15 shows an outline of structure of the above two column bar code. As can be understood from FIG. 15, the two column bar code is constructed of four blocks (A-D).
In this case, recognition flags indicating the upper stage and the lower stage is set in a left data block (A, C), and hence the bar code reader is capable of recognizing whether the block is the upper or lower stage block. No such recognition flag is, however, set in a right data block (B, D), and therefore the bar code reader is incapable of distinguishing between the upper block and the lower block from only the demodulated data corresponding to the right block. Accordingly, if the demodulated data obtained by the block reading method described above are treated also as valid data, there might be a possibility in which the bar code reader mis-recognizes the data in such a case that the bar code of the upper stage is constructed of demodulated data (1) obtained by reading the left data block (A) of the upper stage and demodulated data (4) obtained by reading the right data block (D) of the lower stage. Similarly, there might be the possibility in which the bar code reader mis-recognizes the data in such a case that the bar code of the lower stage is constructed of demodulated data (3) obtained by reading the left data block (C) of the lower stage and demodulated data (2) obtained by reading the right data block (B) of the upper stage. Namely, two combinations of right and left data blocks of the upper and lower stages are possible in the whole two column bar code, however, the bar code reader is incapable of identifying one correct combination among those combinations. Then, if a modulus 10 check with respect to the bar code of each of the upper and lower stages becomes OK by accident, the bar code reader might fall into a misjudgment that the demodulation has been completed in spite of the fact that the demodulated data corresponding to the two column bar code are not reproduced.
Therefore, the prior art bar code reader deals, as valid data, with the demodulated data ((5), (6)) obtained by the continuous reading method, and reproduces the demodulated data corresponding to the whole two column bar code on the basis of only the demodulated data obtained by continuously reading the bar code of the upper stage and the bar code of the lower stage, respectively.
The individual bar codes constituting the two column bar code take a truncate label with a small height in the vertical direction, and therefore a probability that a scan beam travels through each bar code non-stop from one end to the other end, is extremely low. Accordingly, it takes a long time till the demodulated data based on the continuous reading method are obtained, and eventually it takes a long time to reproduce the demodulated data corresponding to the whole two column bar code.

Bar code reader, bar code reading method and computer readable medium


Disclosed embodiments relate to systems and methods structured to mitigate cache conflicts through hardware assisted redirection of pages. In one example, a processor includes a translation cache to store a physical to slice mapping in response to a cache conflict mitigation request corresponding to a page; and a cache controller to determine whether the translation cache comprises the physical to slice mapping; determine whether one of a plurality of slices in a translation table comprises the physical to slice mapping if the translation cache does not comprise the physical to slice mapping, the translation table communicably coupled to a non-volatile memory; and if the translation table does not comprise the physical to slice mapping, redirect the cache conflict mitigation request to the non-volatile memory; and allocate a new physical to slice mapping for the page to one of the plurality of slices in the translation table.

Non-volatile memory based memory systems have a two-level memory (2LM) design in which the Dynamic Random Access Memory (DRAM) acts as a hardware managed limited-associativity cache and the non-volatile memory acts as the operating system managed fully associative main memory. Although the 2LM design can provide speedup for applications that require large memory capacities, the 2LM slows down existing workloads with smaller working-sets due to conflict misses experienced by the hardware managed DRAM cache.
Efficiently mitigating cache conflicts through hardware assisted redirection of pages (HARP) may assist in meeting the needs of processors, for example, performing workloads with large and small working sets or other demands requiring access to memory.

Systems and methods for mitigating dram cache conflicts through hardware assisted redirection of pages (HARP)