Activation of plasminogen to plasma is inhibited by preventing the binding of a receptor binding form of urokinase-type plasminogen activator to a urokinase-type plasminogen activator receptor in a mammal, thereby preventing the urokinase-type plasminogen activator from converting plasminogen into plasmin. DNA fragments which encode for soluble, active fragments of the urokinase-type plasminogen activator are provided.

According to the literature, urokinase-type plasminogen activator (u-PA) has been found in all mammalian species so far investigated. Several findings relate u-PA to tissue degradation and/or cell migration, presumably through a breakdown of the extracellular matrix, caused by plasmin together with other proteolytic enzymes. This relation has been most extensively studied in postlactational involution of the mammary gland and the early phase of trophoblast invasion after implantation of the fertilized egg in the uterus. The hypothesis of a role of u-PA in tissue degradation and cell migration is further supported by the more exact localization made possible by the immunocytochemical findings of u-PA in epithelial cells of involuting mammary glands, in areas with tissue degradation in psoriasis, in association with the release of spermatocytes during spermatogenesis, and in keratinocytes of the epithelial outgrowth during wound healing (see Dan.o slashed. et al., 1988, Gr.o slashed.ndal-Hansen et al., 1988).
It is also conceivable that u-PA plays a role in the degradative phase of inflammation, and there have also been reports that u-PA interferes with the lymphocyte-mediated cytotoxicity against a variety of cells, and a direct role of u-PA in the cytotoxic effect of natural killer cells has been proposed. A role of u-PA has been proposed in angiogenesis and in endothelial cell migration, a process important in tumor growth.
u-PA is produced by many cultured cell types of neoplastic origin. It has been found that explants of tumor tissue released more u-PA than the corresponding normal tissue. u-PA has been identified in extracts from human lung, colon, endometrial, breast, prostate and renal carcinomas, human melanomas, murine mammary tumors, the murine Lewis lung tumor, and in ascites from human peritoneal carcinomatosis. An immunohistochemical study of invasively growing and metastasing Lewis lung carcinomas in mice consistently showed the presence of u-PA, but also a pronounced heterogenecity in the content of u-PA in different parts of the individual tumors. A high u-PA content was found in areas with invasive growth and degradation of surrounding normal tissue, while other areas were devoid of detectable u-PA. The u-PA was located in the cytoplasm of the tumor cells and extracellularly surrounding the tumor cells.
Degradation of the surrounding normal tissue is a central feature of invasiveness of malignant tumors. The constant finding of u-PA in malignant tumors and the findings indicating that u-PA plays a role in tissue degradation in normal physiological events have led to the assumption that u-PA plays a similar role in cancer development. The hypothesis of u-PA playing a role in tissue destruction involves the assumption that plasmin, together with other proteolytic enzymes, degrades the extracellular matrix. It is noteworthy in this context that most components of the extracellular matrix can be degraded by plasmin. These include laminin, fibronectin, proteoglycans, and possibly some types of collagen, but not all. In addition, as originally reported by Vaes and collaborators, plasmin can activate latent collagenases which in turn can degrade the other types of collagen (see Dan.o slashed. et al., 1988).
The majority of the cancer patients in the treatment failure group succumb to the direct effects of the metastases or to complications associated with the treatment of metastases. Therefore, much research has been focused on identifying specific biochemical factors which can be the basis for diagnostic or therapeutic strategies. The extracellular matrix is composed of glycoproteins such as fibronectin and laminin, collagen and proteoglycans. Extracellular matrix becomes focally permeable to cell movement only during tissue healing and remodelling, inflammation, and neoplasia. Liotta (1986) has proposed a three-step hypothesis: The first step is tumor cell attachment via cell surface receptors. The anchored tumor cell next secretes hydrolytic enzymes (or induces host cells to secrete enzymes) which can degrade the matrix locally (including degradation of the attachment components). Matrix lysis most probably takes place in a highly localized region close to the tumor cell surface. The third step is tumor cell locomotion into the region of the matrix modified by proteolysis. Thus, invasion of the matrix is not merely due to passive growth pressure but requires active biochemical mechanisms.
Many research groups have proposed that invasive tumor cells secrete matrix-degrading proteinases. A cascade of proteases including serine proteases and thiol proteases all contribute to facilitating tumor invasion. One of the crucial cascades is the plasminogen activation system. Regulation of the proteolysis can take place at many levels including tumor cell-host cell interactions and protease inhibitors produced by the host or by the tumor cells themselves. Expression of matrix-degrading enzymes is not tumor cell specific. The actively invading tumor cells may merely respond to different regulatory signals compared to their non-invasive counterparts (Liotta, 1986).
The assumption that the plasminogen activation system, through a breakdown of extracellular matrix proteins, plays a role in invasiveness and destruction of normal tissue during growth of malignant tumors is supported by a variety of findings. These include a close correlation between transformation of cells with oncogenic viruses and synthesis of u-PA, the finding that u-PA is involved in tissue destruction in many non-malignant conditions, and the immunohistochemical localization of u-PA in invading areas of tumors (see Dan.o slashed. et al., 1985, Saksela, 1985, for reviews).
Further support for this hypothesis has come from studies with anticatalytic antibodies to u-PA in model systems for invasion and metastasis. Such antibodies were found to decrease metastasis to the lung from a human u-PA producing tumor, HEp-3, transplanted onto the chorioallantoic membrane of chicken embryos (Ossowski and Reich, 1983, Ossowski 1988), penetration of amniotic membranes by B16 melanoma cells (Mignatti et al., 1986), basement membrane invasion by several human and murine cell lines of neoplastic origin (Reich et al., 1988), and formation of lung metastasis after intravenous injection of B16 melanoma cells in mice (Hearing et al., 1988). In some of these studies (Mignatti et al., 1986, Reich et al., 1988), a plasmin-catalyzed activation of procollagenases (see Tryggvason et al., 1987) appeared to be a crucial part of the effect of plasminogen activation.
A requirement for the regulation of a proteolytic cascade system in extracellular processes is the precise localization of its initiation and progression. For example, in the complement and coagulation systems, cellular receptors for various components are known and serve to localize reactions that either promote or terminate the reaction sequence (Muller-Eberhard, 1988, Mann et al., 1988). In the plasminogen activation system, the role of fibrin in the localization of plasminogen activation catalyzed by the tissue-type plasminogen activator (t-PA) is well known (Thorsen et al., 1972, Hoylaerts et al., 1982).
Immunocytochemical studies have suggested that in the invasive areas of tumors, u-PA is located at the membrane of the tumor cells (Skriver et al., 1984), and recent findings indicate that at cell surfaces, u-PA is generally bound to a specific receptor and that this localization may be crucial for the regulation of u-PA catalyzed plasminogen activation in time and space (see Blasi et al., 1987). Preliminary reports suggest that also t-PA may bind to cell surface receptors and retain its enzymatic activity (Beebe, 1987, Barnathan et al., 1988, Hajjar and Nachmann, 1988, Kuiper et al., 1988). This phenomenon, however, awaits further clarification concerning the nature of the binding sites.
Surface receptor for u-PA
The cellular receptor for u-PA (u-PAR) was originally identified in blood monocytes and in the monocyte-like U937 cell line (Vassalli et al., 1985), and its presence has been demonstrated on a variety of cultured cells, including several types of malignant cells (Stoppelli et al., 1985, Vassalli et al., 1985, Plow et al., 1986, Boyd et al., 1988a, Nielsen et al., 1988), human fibroblasts (Bajpai and Baker, 1985), and also in human breast carcinoma tissue (Needham et al., 1987). The receptor binds active 54 kD u-PA, its one-polypeptide chain proenzyme, pro-u-PA (see below), as well as 54 kD u-PA inhibited by the active site reagent DFP, but shows no binding of the low molecular weight (33 kD) form of active u-PA (Vassalli et al., 1985; Cubellis et al., 1986). Thus, binding to the receptor does not require the catalytic site of u-PA, and in agreement with these findings, the binding determinant of u-PA has been identified in the amino-terminal part of the enzyme, in a region which in the primary structure is remote from the catalytic site. The receptor binding domain is located in the 15 kD amino-terminal fragment (ATF, residues 1-135) of the u-PA molecule, more precisely within the cysteine-rich region termed the growth factor region as this region shows homologies to the part of epidermal growth factor (EGF) which is responsible for binding to the EGF receptor. The amino acid residues which appear to be critical for binding are located within the sequence 12-32 (Appella et al., 1987). Synthetic peptides have been constructed that inhibit the binding of very low (100 nM) concentrations. The lack of cross-reactivity between the murine and the human peptides indicates that the binding between u-PA and u-PAR is strongly species specific.
Binding of u-PA to u-PAR is specific in the sense that as yet no other protein has been found to compete for binding to the receptor, though several proteins structurally related to u-PA, including t-PA and plasminogen, have been tested (Stoppelli et al., 1985, Vassalli et al., 1985, Nielsen et al., 1988). Fragments of u-PA containing only the receptor binding domain, e.g. ATF, ensure specificity of the binding to the receptor, since other molecules that might bind u-PA (protease nexin and the specific plasminogen activator inhibitors PAI-1 and PAI-2) recognize the catalytically active region (Stoppelli et al., 1985; Nielsen et al., 1988). PAI-1 is able to form a covalent complex with u-PA but not with pro-u-PA (Andreasen et al., 1986).
The number of receptors reported varies strongly among the cell types studied, from a few thousand molecules per cell on normal monocytes (Miles and Plow, 1987) up to 3.times.10.sup.5 on some colon carcinoma cell lines (Boyd et al., 1988a), and some variation apparently also occurs in the binding affinity, which is in the 0.1-10 nM range (for a review, see Blasi 1988). Further, on certain cell lines the number of receptors can be regulated by the addition of various agents such as phorbol myristate acetate (PMA) in U937 cells (Stoppelli et al., 1985, Nielsen et al., 1988), epidermal growth factor in A431 cells (Blasi et al., 1986) and HeLa cells (Estreicher et al., 1989) and dimethylformamide in colon carcinoma cells (Boyd et al., 1988b). In the first-mentioned case, a large decrease in affinity for the ligand occurs concomitantly with an increase in the number of receptors (Nielsen et al., 1988, Picone et al., 1989).
Preliminary molecular studies on the u-PA receptor have been carried out. A u-PA receptor assay has been developed and an approximately 2200-fold purification has been accomplished, using metabolically labelled material and affinity chromatography with immobilized pro-u-PA (Nielsen et al., 1988). Characterization of the partly purified protein has shown that the receptor is a 55-60 kD glycoprotein, the molecular weight of which is unchanged after cleavage of disulfide bonds, suggesting that it consists of a single polypeptide chain. Until the present invention, nothing was known about the structural properties of the receptor, responsible for binding to the ligand. In the study of Nielsen et al., the purified u-PAR preparation shows essentially one radiolabelled band after SDS-PAGE followed by autoradiography. This analysis, however, does not show the purity of the preparation as it does not detect unlabelled proteins that may be present in an amount that may be higher than that of the u-PAR. Similar considerations hold true for a recent study by Estreicher et al. (1989), in which attempts at purifying u-PAR were done from cells that had been surface-labelled with .sup.125 I. By detergent separation followed by incubation with diisopropylfluorophosphate labelled u-PA (DFP-u-PA) and affinity chromatography with immobilized antibodies to u-PA, a labelled band of approximately 45,000 kD was obtained after SDS-PAGE and autoradiography. It is not clear whether this band represents u-PAR. No cross-linking studies have been performed on the purified preparation, and its apparent molecular weight is distinctly lower than than of u-PAR as reported by Nielsen et al. (1988) and found in the present study (see Example 1). In addition, it cannot be evaluated whether contaminating non-labelled proteins are present, and as only a part of the lane in the SDS-PAGE is shown, even an evaluation of whether contaminating labelled proteins are present is impossible.
Preparation of antibodies to u-PAR has hitherto not been described.
Proenzyme to u-PA (pro-u-PA)
Several studies have indicated that u-PA is released from many types of cultured cells as a single-chain proenzyme with little or no plasminogen activating capacity (Nielsen et al., 1982, Skriver et al., 1982, Eaton et al., 1984, Kasai et al., 1985, Pannell and Gurewich 1987). By limited proteolysis with catalytic amounts of plasmin, this proenzyme can be converted to its active two-chain counterpart. The proenzyme nature of single-chain u-PA is also reflected in the finding that it has essentially no amidolytic activity with synthetic substrates (Wun et al., 1982, Eaton et al., 1984, Lijnen et al., 1986, Stump et al., 1986a, 1986b, Nelles et al., 1987, Pannell and Gurewich 1987), and that it has little or no reactivity with macromolecular inhibitors (Eaton et al., 1984, Vassalli et al., 1985, Andreasen et al., 1986, Stephens et al., 1987) and synthetic inhibitors (Nielsen et al., 1982, Skriver et al., 1982, Wun et al., 1982, Gurewich et al., 1984, Kasai et al., 1985).
This picture of single-chain u-PA as an essentially inactive proenzyme is in contrast to the interpretation reached by some other investigators (Collen et al., 1986, Lijnen et al., 1986, Stump et al., 1986a, 1986b). They concluded that single-chain u-PA from several sources had considerable plasminogen activating capability, and that recombinant single-chain u-PA had an activity that was even higher than that of two-chain u-PA. For these studies, a coupled plasminogen activation assay was used in which the activity of generated plasmin was measured with a chromogenic substrate. Such assays for pro-u-PA are self-activating and are strongly influenced by small amounts of contaminating or generated two-chain u-PA or plasmin. As discussed in detail elsewhere (Petersen et al., 1988), it is therefore possible that the high activity of one-chain u-PA found in these studies was apparent and not due to intrinsic activity of single-chain u-PA. Consistent with this interpretation is a report on a variant of recombinant single-chain u-PA which by site-directed mutagenesis was made partly resistant to plasmin cleavage. This variant of single-chain u-PA had an activity that in coupled assays was 200-fold lower than that of two-chain u-PA (Nelles et al., 1987).
Recent kinetic studies, which included measures to prevent self-activation in the assays for pro-u-PA, have confirmed the low intrinsic activity of pro-u-PA (Ellis et al., 1987, Petersen et al., 1988, Urano et al., 1988). In one study with a highly purified preparation of pro-u-PA from HT-1080 fibrosarcoma cells, it was shown that the pro-u-PA had a capacity for plasminogen activation that was lower than that of a 250-fold lower concentration of two-chain u-PA. It was not possible to decide whether this low activity was intrinsic or due to contamination (Petersen et al., 1988).
In the intact organism, pro-u-PA is the predominant form of u-PA in intracellular stores, and it also constitutes a sizable fraction of the u-PA in extracellular fluids (Skriver et al., 1984, Kielberg et al., 1985). Extracellular activation of pro-u-PA may therefore be a crucial step in the physiological regulation of the u-PA pathway of plasminogen activation. The plasmin-catalyzed activation of pro-u-PA provides a positive feedback mechanism that accelerates and amplifies the effect of activation of a small amount of pro-u-PA. The initiation of the u-PA pathway of plasminogen activation under physiological conditions, however, involves triggering factors that activate pro-u-PA as described herein. Mutants of human single-chain pro-u-PA in which lysine 158 is changed to another amino acid (e.g. Glu or Gly) are not, or are only to a small extent, converted to active two-chain u-PA (Nelles et al., 1987).
u-PA at focal contact sites
At the surface of HT-1080 fibrosarcoma cells and human fibroblasts, u-PA has been found to be unevenly distributed, distinctly located at cell-cell contact sites and at focal contacts that are the sites of closest apposition between the cells and the substratum (Pollanen et al., 1987, 1988, Hebert and Baker 1988). u-PA was not detected in the two other types of cell-substratum contact, i.e. close contacts and fibronexuses, making it an intrinsic component at focal contact sites (Pollanen et al., 1988). u-PA at the focal contact sites is receptor-bound (Hebert and Baker, 1988). The focal contact sites are located at the termini of actin-containing microfilament bundles, the so-called stress fibers or actin cables (Burridge, 1986). These sites contain several structural components (actin, talin) and regulatory factors (the tyrosine kinase protooncogene products P60.sup.src, P120.sup.gag-abl, P90.sup.gag-yes, P80.sup.gag-yes), that are all located on the cytoplasmic side (see Burridge, 1986).
Plasminogen binding sites on cell surfaces
Plasminogen, as well as plasmin, binds to many types of cultured cells, including thrombocytes, endothelial cells and several cell types of neoplastic origin (Miles and Plow, 1985, Hajjar et al., 1986, Plow et al., 1986, Miles and Plow 1987, Burtin and Fondaneche, 1988). The binding is saturable with a rather low affinity for plasminogen (K.sub.D 1 .mu.M). At least in some cell types, binding of plasmin appears to utilize the same site as plasminogen, but the binding parameters for plasmin indicate that more than one type of binding site for plasminogen and plasmin may exist. Thus, on some cell types, plasmin and plasminogen bind with almost equal affinity (Plow et al., 1986), while on others plasmin apparently binds with a higher affinity (K.sub.D 50 nM) than plasminogen (Burtin and Fondaneche, 1988). The binding is inhibited by low amounts of lysine and lysine analogues and appears to involve the kringle structure of the heavy chains of plasminogen and plasmin (Miles et al., 1988).
The binding capacity varies between cell types and in many cell types is quite high (10.sup.5 -10.sup.7 binding sites per cell). The chemical nature of the binding sites are not known. A membrane protein, GPIIb/IIIa, seems to be involved in the binding of plasminogen to thrombocytes (Miles et al., 1986) and, particularly on thrombin-stimulated thrombocytes, also fibrin may be involved in plasminogen binding (Miles et al., 1986). In its purified form, the thrombocyte protein thrombospondin forms complexes (K.sub.D 35 nM) with plasminogen (Silverstein et al., 1984). Also immobilized laminin (Salonen et al., 1984) and fibronectin (Salonen et al., 1985) bind plasminogen (K.sub.D 3 nM and 90 nM, respectively)
Surface plasminogen activation
Some cell types bind both u-PA and plasminogen (Plow et al., 1986, Miles and Plow, 1987, Burtin and Fondaneche, 1988, Ellis et al., 1988). Receptor-bound pro-u-PA can be activated by plasmin (Cubellis et al., 1986) and, at least in part, receptor-bound two-chain u-PA retains its ability to activate plasminogen (Vassalli et al., 1985).
Addition of u-PA and plasminogen to cells holding binding sites for both molecules leads to the occurrence of cell-bound plasmin (Plow et al., 1986, Burtin and Fondaneche, 1988). These studies did not allow a rigorous discrimination between an activation process occurring in solution or between surface-bound reactants.
An interaction between binding sites for u-PA and plasminogen is suggested by the finding that u-PA binding in two cell lines led to an increased binding capacity for plasminogen. Binding of plasminogen in these studies had no effect on the binding capacity for u-PA (Plow et al., 1986). An enhancement of u-PA binding caused by plasminogen was also found by Burtin and Fondaneche (1988) in a cell line of neoplastic origin, even though the plasminogen binding sites demonstrated in the two studies were apparently not identical (see above).
Recently, Ossowski (1988) published findings that the invasive ability of human tumor cells (into modified chick embryo chorioallantoic membranes in an in vivo assay) which have surface u-PA receptors, but which do not produce u-PA, could be augmented by saturating their receptors with exogenous u-PA. This finding, however, is only suggestive (as stated by the author) and it does not demonstrate that binding to the receptor per se is necessary. It is possible that the u-PA added to the cells was carried to the site of invasiveness because of receptor binding, but released from the receptor before exerting its activation. In addition, this study was carried out with two-chain u-PA and therefore does not simulate endogenous u-PA of the single-chain form. In the study of Ossowski, it was also found that an increased production of mouse u-PA in human cells transfected with mouse u-PA cDNA under the control of a human heat shock promoter did not increase invasiveness. Mouse u-PA does not bind to human u-PAR, but the published data cannot be taken as a proof that this lack of effect of mouse u-PA is due to this lack of receptor binding because several other explanations are possible, e.g. 1) that the mouse u-PA does not activate chicken plasminogen as efficiently as human u-PA, 2) that in this system there are lacking mechanisms of converting one-chain mouse u-PA to the two-chain form, 3) that the heat shock in itself decreases the ability of the cells to invade, 4) that the heat shock treatment does not increase the production of mouse u-PA when it is followed by implantation that changes the microenvironments of the cells. No attempts were made in this study to investigate the effect on invasion of displacement of u-PA from its receptor.
Ellis et al. (1989) recently published evidence indicating that the reactions leading to plasminogen activation can take place when single-chain u-PA and plasminogen are added to U937 cells, and that they occur more efficiently when both plasminogen and pro-u-PA are bound to the surface. This experiment, however, was performed in the absence of serum, i.e. under conditions where the plasminogen activation with the preparations used by Ellis et al. will also take place in solution (cf. Ellis et al., 1987), and these studies do not exclude the possibility that one or more of the processes involved (e.g. the plasminogen activation catalyzed by two-chain u-PA) actually occurred when the u-PA was not receptor-bound. Moreover, these studies used a purified preparation of single-chain u-PA that has a catalytic activity considerably higher than that found for single-chain u-PA by other groups (Pannell and Gurewich, 1987; Urano et al., 1988; Petersen et al., 1988). Ellis' preparation may therefore be contaminated with two-chain u-PA and thus be distinctly different from the endogenous single-chain u-PA produced by cells in situ. In the experiments according to Ellis et al., 1989, binding of the added single-chain u-PA to the receptor was prevented by preincubation of the cells with the amino-terminal fragment of u-PA. These experiments do not, therefore, as do the following examples, demonstrate displacement of endogenously produced u-PA, a prerequisite for any therapeutic use of this approach.

Vectors and methods for recombinant production of uPA-binding fragments of the human urokinase-type plasminogen receptor (uPAR)


A system and methods of securely storing biological samples prior to the secure transportation of the biological and/or biohazardous specimens to a depository where the system can determine the locations and pickup times of the couriers transporting the enclosures from the storage unit to the depository.

1. Field of the Invention
Among other things, the present invention includes a system and methods and devices for securely storing biological samples prior to the secure transportation of the biological and/or biohazardous specimens to a depository. Within the scope of the present system and method, enclosures for securely enclosing the biological and/or biohazardous specimens in accordance with national, state and local safe handling standards for said biological specimens are supplied to specimen handlers. The current system and methods are capable of determining the locations and pickup times of the couriers transporting the enclosures from the storage unit to the depository.
2. Description of the Previous Art
1) U.S. Pat. No. 4,576,281-Kirksey enables a disposable syringe needle separation and storage box. The Description of the Preferred Embodiment, in part, reads, "In that respect, the present invention is directed to a separated syringe needle storage enclosure or box for facilitating the disposal of such needles after separation from the syringe barrel with such enclosure having means for facilitating the separation thereof. Further, it should be appreciated that certain components of the enclosure and disposal elements have counterparts in the referred to prior art apparatus. The improved, vertically upright, syringe needle storage enclosure box for facilitating the disposal of such separated needles, is indicated generally at 22. The vertically upright, outer box may be formed of steel although the illustrated box is of wood. In that respect, the enclosure 22 includes a top wall 24, a bottom wall 26, laterally opposed sidewalls 28 and 30, a rear wall 32 and a front opening door 34. The door 34 is mounted to the right sidewall 30 by way of a continuous hinge 36, the hinge 36 being appropriately screwed to the end of sidewall 30 and to the end of the door 34 by means of wood screws 38. Rubber grommets at 40 are fixedly mounted to the lower surface of the bottom wall 26 to permit the enclosure 22 to stand upright on the surface of a desk, cabinet or the like. Enclosure 22 is essentially identical to the cited prior art outer enclosure, and may be made of steel or other sheet metal in the manner of the prior art structures, if desired.
Additionally, the top wall 24 is provided with a circular opening or hole 42. Unlike the prior art, instead of a basin or dish bearing a plastic bag functioning as the inner container for receiving separated disposable syringe needles or needle assemblies, which are gravity dropped into such container, the present invention employs a molded plastic jar indicated generally at 44 which is of a diameter approximating the width of the enclosure 22, and enclosure 22 may be square in cross-section. Further, the height of the jar 44 is such that it does not reach to the top wall 24 of the outer enclosure 22. The plastic jar 44 is necked down at its upper end as at 44a, terminating in a reduced diameter throat or rim 44b, which throat or rim 44b is threaded on its outer surface as at 46. As such, when a sufficient quantity of needle assemblies 16 accumulate within the jar 44, the door 34 may be unlocked, the jar 44 removed, and a cap (not shown) threaded to the top of the jar, The cap may be metal or plastic but should be sufficiently strong to prevent the sharp point 20a of the metal needle 20 from penetrating through the wall of the plastic jar 44 or its cap (not shown)."
Based on the current record, among other things, it does not appear that Kirksey teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Kirksey does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
2) U.S. Pat. No. 5,818,336—Varga, et al. enables a drop box inventory monitoring and control system. The Description of the Preferred Embodiment, in part, reads, "Referring now to the drawings, wherein like characters designate like or corresponding parts throughout the several views, there is shown in FIG. 1 an inventory monitoring and control system 100 according a preferred embodiment of the present invention. System 100 includes a drop sensor 200 cooperatively coupled for communication through a network modem 300. The drop sensor 200, network modem 300 and a door switch 400 are each powered for operation by an AC/DC power supply 500. Any network, such as a network radio modem or a public service telephone modem, may be used as network modem 300 In such a manner, system 100 can monitor and control inventories contained, for example, within a drop box 800, 850 as shown in FIGS. 2 and 3. In a first embodiment, the drop sensor 200 passively detects the passage of packages into the drop box 800, 850, as more fully described below. In a second embodiment, the drop sensor 200 actively detects packages, such as the physical detection of packages by switches, such as a paddle switch. The door switch 400 senses that the courier has opened a locked access door (not shown) of drop box 800, 850 to pick-up packages and commands the network modem 300 to transmit a message that the packages have been picked up.
With reference first to the embodiment shown in FIG. 2, the drop sensor 200 is positioned within drop box 800 of the type having a first door 810 through which a patron deposits a package P. Attached to the first door 810 is an extension 820 which generally propels the package P into a downward trajectory within the drop box 800. Drop sensor 200 is, thus, positioned within the drop box 800 such that its sensing field F is generally parallel to the floor 830 of drop box 800. In a conventional manner, drop box 800 includes a second, courier door 840 for removal of the packages P deposited therein.
Drop box 850, as shown in FIG. 3, also includes a first door 810 which is adapted for receiving packages P deposited by a patron, and a second door 840 which permits the courier to remove those packages P deposited within drop box 850. It should be readily apparent from FIG. 3 that the first door 810 of drop box 850 does not include an extension 820 as does its counterpart drop box 800. In such cases, packages P may not break the sensing field F of the drop sensor 200 if positioned as shown in FIG. 2. Accordingly, the drop sensor 200 shown in FIG. 3 is positioned optimally to project its sensing field F downwardly across the drop box 850 so that, in the unlikely event that a package P falls in a generally parallel position with respect to the floor 830, such package P will nevertheless be sensed by the drop sensor 200. In accordance with a presently preferred embodiment of the invention, drop sensors 200 deployed within drop boxes 850 of the type shown in FIG. 3 should be positioned such that their sensing field F is approximately 35° below a line which is parallel to the floor 830 of those drop boxes 850."
Based on the current record, among other things, it does not appear that Varga teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Varga does not teach or disclose the utilization of a storage unit communications module capable of intercommunication with multiple storage unit hardware components, where the communications module can send/receive data with a network.
3) U.S. Pat. No. 7,640,769—Clark enables a portable self-contained storage apparatus for biologicals. The Detailed Description of Invention, in part, reads, "FIG. 2 is a front cross-sectional view of a first embodiment of the invention (vertical section II-II′ or VI-VP—see FIGS. 1 and 5). Cabinet 1 contains a thermally-insulated box 20, which is subdivided into an upper (freezer) compartment 21 and a main (chilled) compartment 22. The two subdivisions are insulated from each other by an insulated barrier 24. Below the box 20 is a power compartment 25 containing a battery pack 26 and a compressor 27 along with electronic regulating circuitry. High-efficiency insulation 204 (e.g., rigid foam) is interposed between box 20, cabinet 1, and power compartment 25. Also visible in this view are lifting handles 6 and feet 7. Both the freezer compartment 21 and the chilled compartment 22 contain one or more tray support racks 28. (In the depicted embodiment, the freezer compartment 21 contains two such racks, and the chilled compartment 22 contains four such racks). Each rack 28 supports a plurality of elongate trays 201 (in the depicted embodiment, five) extending from the front of the box to the back. The trays 201 are of a width to accommodate the long dimension of a standard vial box 203 (shown in dashed lines to indicate environmental structure)."
Based on the current record, among other things, it does not appear that Clark teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Clark does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
4) US Patent 20020125998—Petite, et al., discloses a system and method for monitoring and controlling remote devices. The Summary of the Invention, in part, reads, "[0014] In accordance with a broad aspect of the invention, a system is provided having one or more sensors to be read and/or actuators to be controlled remotely, ultimately through a computer on the Internet. The sensors and/or actuators are interfaced with wireless transceivers that transmit and/or receive data to and from the Internet. In this regard, additional wireless transceivers may relay information between the transceivers disposed in connection with the sensors and actuators and a gateway to the Internet. It should be appreciated that, a portion of the information communicated includes data that uniquely identifies the sensors and/or actuators.
[0015] In accordance with one aspect of the invention, a system is configured to monitor and report system parameters. The system is implemented by using a plurality of wireless transceivers. At least one wireless transceiver is interfaced with a sensor, transducer, actuator or some other device associated with the application parameter of interest. In this regard, the term "parameter" is broadly construed and may include, but is not limited to, a system alarm condition, a system process variable, an operational condition, etc. The system also includes a plurality of transceivers that act as signal repeaters that are dispersed throughout the nearby geographic region at defined locations. By defined locations, it is meant only that the location of each transceiver is known to a central computer. The central computer may be informed of transceiver physical locations after permanent installation, as the installation location of the transceivers is not limited. Each transceiver that serves to repeat a previously generated data signal may be further integrated with its own unique sensor or a sensor actuator combination as required. Additional transceivers may be configured as stand-alone devices that serve to simply receive, format, and further transmit system data signals. Further, the system includes a local data formatter that is configured to receive information communicated from the transceivers, format the data, and forward the data via the gateway to one or more servers interconnected with the WAN. The server further includes means for evaluating the received information and identifying the system parameter and the originating location of the parameter. The server also includes means for updating a database or further processing the reported parameters.
[0016] Consistent with the broader concepts of the invention, the "means" for evaluating the received information and the "means" for reporting system parameters are not limited to a particular embodiment or configuration. Preferably, these "means" will be implemented in software that is executed by a processor within a server integrated with the Internet. However, dedicated WANs or Intranets are suitable backbones for implementing defined system data transfer functions consistent with the invention."
Based on the current record, among other things, it does not appear that Petite teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Petite does not teach or disclose the utilization of a storage unit communications module capable of intercommunication with multiple storage unit hardware components, where the communications module can send/receive data with a network.
5) U.S. Pat. No. 6,299,061—Henson enables a security mailbox. The Description of the Preferred Embodiment, in part, reads, "As a first embodiment of the security mailbox 10, the means for closing access to the compartment 11 through the slot 18 includes a revolving door member 22 conventionally mounted inside the container 11 and having a shaft 23 rotatably and conventionally mounted to the side walls 13,14 proximate to the slot 18, and further has a plurality of door members 24-27 having ends securely and conventionally attached to the shaft 23 for rotation therewith and extending radially therefrom. The door members 24-27 are adapted to close access to the compartment 11 between themselves and the front wall 17 at the slot 18, one of the door members 27 having an outer end which has a weighted member 31 securely attached thereto for placing the door members 24-27 in a closed position."
Based on the current record, among other things, it does not appear that Henson teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for said biological specimens. Further, Henson does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
6) US Patent 120578—Farrington enables an improvement in letter boxes. Page 1 of Farrington reads, "One edge of each of these shelves is rigidly secure, longitudinally, to a horizontal shaft, E, the other edge is free to move by the weight of the newspaper when dropped upon it. F is a curved guard, fitted to the line of motion of the outer edges of the revolving shelves . . . upon which said guard the paper are deposited by the revolving shelves . . . "
Based on the current record, among other things, it does not appear that Farrington teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Farrington does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
7) U.S. Pat. No. 6,742,703—Esakov, et al., enables a mail collection box. The Detailed Description of the Invention, in part, reads, "A deposit door 120 may be supported by housing 102, for example pivotally supported by one or more hinges mounted to housing 102, so that the deposit door is moveable from a deposit door open position 122 (FIGS. 1, 4-5, 8), which provides access to the housing interior space 104 through mail deposit opening 118, and a deposit door closed position 124 (FIGS. 2-4), in which the perimeter 126 of deposit door 120 engages housing 102. Deposit door 120 may include a deposit opening gasket 176 proximate perimeter 126—and/or housing 102 may include deposit opening gasket 176 surrounding mail deposit opening 118—to facilitate a seal between the deposit door and the housing in the deposit door closed position 124. The quality and type of seal may be that of any of the seals described below. Deposit door locking mechanism 142 is positioned to interact between the deposit door 120 and housing 102 so that the deposit door 102 may be locked in the deposit door closed position 124. Portions of locking mechanism 142 may be mounted to housing 102, deposit door 120, or both. Suitable locking mechanisms are known in the art."
Based on the current record, among other things, it does not appear that Esakov teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Esakov does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
8) US Patent 20060113368—Dudley discloses a theft preventative mailbox having remote unlocking activation mechanism. The Detailed Description of Preferred Embodiments, in part, reads, "The trap door mechanism 203 can be implemented in a variety of ways. The trap door mechanism 203 can be in a default closed position so that outgoing mail can be placed into the main compartment 101 at any time. Alternatively, the trap door mechanism 203 can have the trap door(s) 201 in the default open position, with the trap door(s) 201 movable to the closed position by the addressee if the addressee desires to place outgoing mail in the main compartment 101. In either case, the trap door(s) 201 are adaptable to retain outgoing mail in the main compartment 101 and permit incoming mail to eventually fall into the secure drop box compartment 202. Although two trap door mechanisms are described below, there are a variety of different trap door mechanisms that can be used in the various embodiments of the present invention, all of which are covered by this present invention. The trap door(s) 201 can be comprised of a single door or it can be comprised of multiple trap doors, such as two halves that open in the center, such trap door(s) 201 and being hinge-ably coupled proximate the top of the secure drop box compartment 202. A first trap door mechanism 203 includes a release button or disc that is manually operated. The default position of the trap door(s) 201 is the closed position. When the postal employee inserts any new mail into the main compartment 101 and presses the release button or disc, the trap door(s) open and drop the incoming mail into the secure drop box compartment 202. The release button or disc and trap door(s) 201 can further be coupled to a mechanical or electronic time delay mechanism such that after the mail is deposited, the trap door(s) 201 automatically close after a predetermined amount of time. When the postal employee closes the front mailbox door 102, the locking and unlocking mechanism 103 is adapted to automatically lock.
A second trap door mechanism can be coupled to the unlocking activation means 104 such that after a certain amount of time after the unlocking activation means 104 is activated by the postal employee using a specially encoded signal, and the front mailbox door 102 is opened, the trap door(s) 201 automatically open, thus dropping any items placed in the main compartment 101 into the secure drop box compartment 202. The default position of the trap door(s) 201 that use this second trap door mechanism is the closed position. After a further amount of time as determined by a mechanical or electronic timing mechanism, the trap door(s) 201 close.
Once packages and mail have fallen into the secure drop box compartment 202, the trap door(s) 201 return to the closed position leaving the incoming mail in the secure drop box compartment 202 until the addressee retrieves the mail by opening at least one drop box door 204 located on the backside of the mailbox.
The drop box door(s) 204 is hinge-ably connected to the secure drop box compartment 202 and is adapted to allow access into the secure drop box compartment 202. The drop box door(s) 204 includes a drop box door lock mechanism 205 that is secured by a key lock or is integral with the card key system 400, FOB system 500 or RFID system 600 unlocking activation mechanism 104 described herein. The drop box door(s) 204 can only be unlocked using the addressee's specific key, card key, FOB transmitter, RFID reader or override key. In the card key system, a contact reader can be used which comprises a slot located on the backside of the mailbox. When the addressee closes the drop box door(s) 204, the drop box lock mechanism 205 is adapted to automatically lock.
In a first embodiment of the present invention, a curbside mailbox with an unlocking activation mechanism 104 that is activated using a card key system 400 is provided. As seen in FIG. 4, the unlocking activation mechanism 104 can be comprised of a card key system 400 having a master coded card 401A, a subordinate card 401 B retained by the addressee and a card reader 402, the card reader 402 being located at the mailbox and being coupled to the locking and unlocking mechanism 103. The card reader 402 would have a power source which can be the AC mains, a non-rechargeable or rechargeable DC battery source or a DC solar power source which is operable to power the card reader 402 directly or to recharge the rechargeable batteries. Card key systems can comprise a variety of technologies, such as contact, contactless (proximity), passive, active, magnetic, electronic, sonar and optical. Each of such implementation technologies is included within the purview of this invention."
Based on the current record, among other things, it does not appear that Dudley teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Dudley does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
9) US Published Patent Application 20080067227—Poss, et al. discloses an electrically-powered programmable package deposit enclosure. The Detailed Description, in part, reads, "FIG. 1 provides a perspective view detailing outer container 24, insertion door 22, Envelopes and Mailers door 21, Envelopes and Mailers shelves 20, Package Removal Door 26, showing the relationship between the doors and compartments according to this embodiment of the present invention. A photovoltaic (PV) cell array 32 is mounted on top of the unit, covering part of it. In one embodiment, cells 32 produce enough power for the average number of 15 data logging events and wireless data transmissions per day. The battery 36, shown in FIG. 2, has enough energy storage to provide for usage through several weeks of intermittent sunlight, to provide adequate reserve power for periods of bad weather. The cells are wired to the energy storage system, which stores power to drive the sensors and Programmable Logic Controller PLC 44. Status indicator lamps 60, FIGS. 3 and 4, provide visual means of displaying information such as a system malfunction or to indicate the level of bin capacity used and available or other notices, such as "low battery," "bin collected," "envelopes empty," "device broken," or other messages. An insertion door 22 acts to prevent unauthorized use because it can lock out the user from the insertion door. In one embodiment, the insertion door lock is opened when it receives a message from PLC 44. In this embodiment, PLC will send a message to open the insertion door when it receives input from a Barcode Scanner 68, FIG. 4. This input may be given when, for example, a package with the appropriate Bar Code is placed in front of the scanner and the "ON Button," 67 in FIG. 4, is pressed by the user. In an alternative embodiment, the device is turned on when the Insertion Door 22 is opened or when the ON Button is pressed."
Based on the current record, among other things, it does not appear that Poss teaches or discloses an ultraviolet light irradiating, for a predetermined time, the storage unit to reduce microbial growth.
10) U.S. Pat. No. 5,774,053—Porter enables a storage device for the delivery and pickup of goods. The Detailed Description of the Preferred Embodiments, in part, reads, "As best illustrated in FIG. 3, the enclosure 14 includes a front hinged door 18 for permitting access to the front of the enclosure 14 and may include a rear hinged door 20 for permitting the homeowner to retrieve goods from or place goods in the rear of the enclosure 14. In the first embodiment of the storage device 10 illustrated in FIG. 1, the rear hinged door 20 may extend through an exterior wall of the home 12 so that the homeowner can access the storage device 10 while inside the home 12."
Based on the current record, among other things, it does not appear that Porter teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Porter does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
11) U.S. Pat. No. 6,967,575—Dohrmann, et al. enables methods and apparatus for unattended pickups and deliveries. The Detailed Description, in part, reads, "FIG. 2 is a perspective view of a secure pickup and delivery container 202, in accordance with one embodiment of the invention. Secure container 202 includes a lockable door 204 that can include a handle 206. In one embodiment door 204 is coupled to secure container 202 by means of suitable hinges 208. Door 204 allows access to an interior part of the secure container 202. Other types of doors besides hinged doors could be used, such as sliding, shuttered, or revolving doors. Secure container 202 includes an access element 210 that can be implemented in a variety of ways, such as with a token or card slot 212 or a keypad, as described further below. A delivery person must present access request information to access element 210 that matches stored access privilege information in order to gain physical access to secure container 202."
Based on the current record, among other things, it does not appear that Dohrmann teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Dohrmann does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.
12) U.S. Pat. No. 7,256,691—Awobue enables a smart mailbox. The Brief Description of the Preferred Embodiments, in part, reads, "The mailbox 10 is provided with an upper lid 18 movably attached through the hinges 102 (visible in FIG. 2) to the back panel of the mailbox. An upper keyhole 20 positioned on a front panel of the lid 18. A detailed diagram of the keyhole 20 and associated locking system is shown in FIG. 4. The mailbox further comprises a keypad 22 for electronically locking and opening the door, a front door keyhole 24 providing options for mechanically and electronically locking and opening door, and indicators 26 for indicating receipt of mail in the box 12."
Based on the current record, among other things, it does not appear that Awobue teaches or discloses a storage unit with a lower section and an upper section, where the upper section includes a deposit-only gate for receiving deposits of enclosures that enclose biological specimens such that after deposit onto and closure of the deposit-only gate the enclosures drop into a portable insulated holder positioned in the lower section of the storage unit until removal of the enclosures is authorized, and where the deposited enclosures are in accordance with national, state and local safe handling standards for the biological specimens. Further, Awobue does not teach or disclose the utilization of an operations center, a network or a storage unit with electronic components capable of intercommunication with the operations center via the network.

System and methods of preserving integrity and securely transporting biological specimens to a depository and devices for securely storing biological specimens


Various arrangements for determining a location of a base station without timing synchronization are presented. A mobile device may determine that it is moving faster than a threshold velocity. The mobile device may capture a first unsynchronized time of arrival (TOA) measurement and determine an associated first location, wherein the first unsynchronized TOA measurement is based on a first unsynchronized timing measurement of a first received reference signal. The mobile device may capture a second unsynchronized TOA measurement and determine an associated second location, wherein the second unsynchronized TOA measurement is based on a second unsynchronized timing measurement of a second received reference signal. Based on the mobile device moving faster than the threshold velocity, the first location, the second location, the first unsynchronized TOA measurement, and the second unsynchronized TOA measurement may be used for determining the location of the base station.

The usefulness of accurate positioning is well known. A typical form of location determination involves using a satellite-based positioning system, such as the Global Positioning System (GPS). While satellite-based positioning systems can be highly accurate, in some situations it may not be possible to receive signals from a sufficient number of satellite-based positioning system satellites to perform an accurate location determination of a wireless mobile device. For example, in a city dense with buildings, other positioning methods may be more accurate. Range-based positioning systems may be useful for mobile wireless devices that communicate with cellular network base stations. However, in order to determine a location based on ranging information with one or more base stations, the base station's location needs to be known. Many wireless service providers do not make the locations of their base stations publicly available, therefore the location of these base stations may need to be determined Such determination of the location of a base station may be referred to as "reverse positioning."

Method and apparatus for asynchrosous positioning of wireless base stations


A process for the formation of pellets containing an ultra hard (superhard) core coated with an encapsulating material includes utilizing a shovel rotor in combination with a rotating pan and/or a fluidized bed apparatus in sequence. The process includes providing a source of superhard material, providing a mixture comprising a binder, a solvent or fluid medium and the intended coating or encapsulating layer, and combining the superhard material and the mixture in a shovel rotor. The rotor of the shovel rotor is rotated at a velocity such that the superhard material is encapsulated by the mixture to form pellets. The pellets are introduced into a rotating vessel or fluidized bed granulating apparatus, and the pellets are contacted with encapsulating material to form pellets of greater mass than the pellets introduced into the vessel.

U.S. Pat. No. 4,770,907 (Kimura, Fuji Paudal, 1988), U.S. Pat. No. 5,143,523 (Matarrese, GE, 1992) and U.S. Pat. No. 5,405,573 (Clark et al., GE, 1995) all describe the use of fluidised beds to coat particles. In this method, ultra hard cores, e.g. diamond seeds, are suspended in a flow of gas within a chamber, into which a fine suspension of binder, solvent and particulate material (e.g. metal powder, the encapsulating material) is sprayed. Alternatively, the binder-solvent may be sprayed with separate powder addition. The emergent pellets are built up in volume proportional (non-linearly) to the residence time spent in the chamber. The advantage of this process is that the fluid bed allows a good separation of the core seeds and thereby ensures that a single core (diamond seed) is contained in each pellet while depositing coating material at a reasonable rate. The disadvantage of this technique is that the maximum deposition rate is relatively slow and when using a high density particulate coating material e.g. Mb, W and WC, and the increasing mass of the pellets presents difficulties in terms of the capabilities of the equipment to maintain the suspension. This can be addressed by increasing the capacity of the equipment but this is costly and impacts on the commercial viability of producing commercial volumes of material. In addition, fluid-bed type coatings are not ideal when coating with a mix of powders of different densities and particle shapes and sizes as the material has a tendency to segregate in the fluid bed and the intended composition may not be uniformly achieved.
The "rotating pan" method as described in GB1014295 (Norton, 1965) and EP0012631 (Tomlinson, DeBeers, 1983) involves introducing the ultra hard core material, e.g. diamond seeds, into either a rotating inclined pan, a drum or any other rotating vessel, where the pellet can be built up by 1) spraying a slurry containing metal powder, binder and solvent (encapsulating material) over the rotating diamond seeds and/or 2) the binder and solvent is/are sprayed separately and the metal powder then "sprinkled" over the rotating diamond seeds. Rotation of the pan separates the coated diamond seeds (emergent pellets) and allows time for removal of the solvent from the sprayed material to form a concentric jacket of encapsulating material which increases in volume as the process proceeds. This technique is efficient in terms of depositing encapsulating material and thus building up the pellet mass quickly. However, the difficulty with this method is that it is susceptible to agglomeration of the cores and/or early pellets in the initial stages of the process. Deposition rates must be very slow to avoid agglomeration and this increases the overall processing time and reduces the throughput of the process. Agglomeration reduces in severity after the emergent pellet has attained a critical size. The consequence of the agglomeration is that the final pellets may have significant size distribution and may contain more than one core per pellet. This contributes to increased process time and cost.
ZA2006/06674 (Flynn et al., E6, 2006) partly over come this problem by adopting a dual stage process for the rapid formation of pellets. Essentially the solution to the problems described above is to combine the two techniques known in the art into a single process design. As such, the initial stages of the process involve a fluid bed approach to maximise the yield of pellets containing one core particle only e.g. diamond seeds. The pellets may be built up to a critical size volume (Vcrit) whilst remaining in a fluid suspension. When the pellets attain this critical size, the pellets are transferred to a rotating pan where the pellets form the (sub) core of the final pellet process. The pellets so produced have a volume significantly greater than the pellets as introduced and the risk of agglomeration is much reduced as the layer on the surface absorbs the spray more quickly and thus deposition rates may be increased. In addition, the weightier particles are less likely to be held together by surface tension of the spray. However, combining both techniques requires two sets of capital equipment and downtime in terms of transferring material from one machine to the other.
Therefore, there is a need to develop a technique which would overcome the technical difficulties outlined above and provide a more cost efficient solution to producing encapsulated abrasives. Preferably such a technique provides an advantage when used in combination with either 'pan rotating' or 'fluidised' route.
The contents of applicant's South African patent application no. 2007/06077 filed on 23 Jul. 2007 and all applications claiming priority therefrom are herein incorporated by reference.

Method for manufacturing encapsulated superhard material


An exhaust-gas cleaning device for an internal combustion engine has a catalytic converter arrangement. The catalytic converter arrangement includes, along the flow direction, a plurality of catalytically active regions which are, in particular, independent of one another. A heat dissipation device includes a heat absorbing device and one heat sink. The heat absorbing device is provided between at least two catalytically active regions. The heat absorbing device is connected to the heat sink. The exhaust-gas cleaning device includes a plurality of catalytic converter arrangements, an, in particular tubular, inner housing with a housing cover, and an outer housing which encloses the inner housing. The inner housing has a longitudinal axis, and the catalytic converter arrangements are arranged separately from one another on the housing cover of the inner housing.

1. Field of the Invention
The invention relates to an exhaust-gas cleaning device for an internal combustion engine, in particular for a stationary gas engine, with at least one catalytic converter arrangement, wherein exhaust gas from the internal combustion engine can flow in a flow direction through the catalytic converter arrangement, and comprises, along the flow direction, a plurality of catalytically active regions that are in particular independent of one another, wherein a heat dissipation device is provided which comprises at least one heat absorbing device and a heat sink, wherein the at least one heat absorbing device is provided between at least two catalytically active regions, the at least one heat absorbing device being in contact with the heat sink.
2. Description of Related Art
Modern high-performance lean burn engines, in particular stationary gas engines, have very high power densities and efficiencies with relatively low manufacturing and maintenance costs. For this reason lean gas engine units are among the most economical energy generation technologies on the market. Compared to diesel engines the emissions of pollutants and toxic substances is also very low, in which connection NOx emissions can be reduced to very low values by internal engine measures and CO and formaldehyde emissions by exhaust gas catalytic measures. Natural gas-operated lean engines on the other hand have relatively high emission values in terms of unburned hydrocarbons, in particular methane (CH4). In contrast to CO, conventional exhaust gas cleaning devices can convert only very small amounts of CH4. In order to achieve sufficiently high conversion rates, in addition to special formulations for the active surface of the catalytic converter arrangements of exhaust gas cleaning devices temperatures of at least about 600° C. are also necessary.
Since the exhaust gas temperature of modern highly supercharged lean burn gas engines after an exhaust gas turbocharger is only about 300-400° C., an exhaust gas cleaning device that is effective for CH4 cannot be used after the exhaust gas turbocharger. In order to achieve a corresponding reaction for the conversion of unburned hydrocarbons in an exhaust gas cleaning device of a highly supercharged lean burn gas engine, the exhaust gas cleaning device must therefore be arranged in front of an exhaust gas turbine. In this way it can be ensured that the temperature necessary for this chemical reaction is achieved.
In the conversion of the unburned hydrocarbons part of the chemical energy contained therein is converted by the chemical reaction into heat. Since the reaction rate in the exhaust gas cleaning device is a function of the temperature, the conversion takes place more rapidly and completely the higher the temperature. Therefore as soon as the concentration of unburned hydrocarbons exceeds a certain value, the rise in temperature on account of the reaction leads to an increase in the reaction rate and thus to a self-sustaining process. The temperature continues to increase until all the unburned hydrocarbons contained in the exhaust gas are completely converted in a very short stretch. With correspondingly increased concentrations of unburned hydrocarbons there is consequently a localized thermal overloading of the catalyst support or the carrier material of the catalytic converter arrangements even melts. This effect is further intensified in the case where the exhaust gas cleaning device is installed in front of an exhaust gas turbine, since the exhaust gas is under a pressure of about 3-4 bars and the energy density of the chemical energy contained in the unburned hydrocarbons is therefore correspondingly increased. One of the most serious problems in this connection is the thermal destruction of regions of an exhaust gas cleaning device due to too high concentrations of unburned hydrocarbons, such as occur for example with ignition failures or misfiring. In principle however it is virtually impossible to avoid ignition failures in gas engines.
As a countermeasure against such an intensified corrosion of a catalyst support body on account of highly exothermic reactions caused by unburned hydrocarbons, DE 100 46 278 A1 for example shows a catalyst support body that has two different zones in the flow direction of the exhaust gas, wherein the first zone has a higher surface specific heat capacity than the second zone. In this way a relatively large amount of thermal energy is extracted from the exhaust gas in the first zone, whereby an intensified corrosion in the second zone can be counteracted. The disadvantage in this case however is that the heat absorption takes place through the catalyst support body itself. Since the heat capacity and the thermal conductivity of the catalyst support body are limited, there is therefore only a slight dissipation of the generated heat from the catalytic converter arrangement to the surrounding structure.
Exhaust gas cleaning devices with respectively a catalytic converter arrangement are also known from U.S. Pat. No. 5,474,745 A and EP 1 111 212 A2, which comprise cooling devices for dissipating heat from the catalytic converter arrangement.

Catalytic converter arrangement for an exhaust-gas cleaning device of an internal combustion engine


In an integrated circuit for electrically permanently program information in the integrated circuit a current is driven through an NPN-transistor. The integrated circuit has a first current feeding circuit having an output connected via a resistor to the NPN-transistor, a feeding point connectable to a feeding voltage, and a control input for controlling the programming of the NPN-transistor. The integrated circuit also has at least a second current feeding circuit having an output connected to the feeding point for the first current feeding circuit, a feeding point connectable to a feeding voltage, and a control input for controlling the programming of the NPN-transistor. Finally the integrated circuit has a first voltage level controller provided to split the voltage needed to program the NPN-transistor over the first and at least second current feeding circuit.

One way to permanently and electrically program bi-polar integrated circuits is to use so-called Zener-zap of NPN-transistors. The NPN-transistor can be programmed by forcing such a strong current through the transistor so that aluminium from the interconnect pattern migrates into the silicon and forms a short-circuit between the base and the emitter.
The NPN-transistor can be used as a logical one or a logical zero when it is zapped, depending on the connected logic. Thus, a sequence of initially non-zapped NPN-transistors can be used to represent a sequence of binary numbers, representing a digital number. This sequence may then be altered, by zapping selected binary numbers in the sequence, to change the digital number represented by said sequence of NPN-transistors.
As an example, the operating point for a power transistor may initially be set to a predetermined value in an integrated circuit. After testing and tuning it is possible that the operating point should be changed slightly to achieve better performance. If the predetermined value for the operating point is set in the integrated circuit using NPN-transistors this value may be changed later, after tuning, by zapping carefully selected NPN-transistors.
It is possible to connect the NPN-transistor in two different ways to produce a Zener diode, namely with the collector connected either to the base or to the emitter. The collector-base connection requires less voltage to zap the Zener diode but the consumed current is considerably larger. If used as a memory function the collector-emitter connection is preferable since less current is consumed. If larger memories should be programmed the current requirements could otherwise be excessive. This alternative has, however, the draw-back that a larger voltage is required.
For circuits involving many components small dimensions are required. These components, having small dimensions, are less tolerant to high voltages. That is, they cannot sustain high voltage without being damaged. It would be preferable to be able to program many NPN-transistors simultaneously also in circuits involving many, and thus small, components, which are incapable of sustaining high voltages.


The present invention relates to cupredoxin and cytochrome and their use, separately or together, to inhibit the spread of parasitemia in mammalian red blood cells and other tissues infected by the malaria parasite, and in particular the parasitemia of human red blood cells by P. falciparum. The invention provides isolated peptides that are variants, derivatives or structural equivalents of cupredoxins or cytochrome c, and compositions comprising cupredoxins and/or cytochrome c, or variants, derivatives or structural equivalents thereof, that are useful for treating or preventing malaria infection in mammals. Further, the invention provides methods to treat mammalian patients to prevent or inhibit the growth of malarial infection in mammals. The invention also provides methods to prevent the growth of malaria infection in insect vectors.

About one quarter of the world's population is exposed to the risk of malaria and more than a million people die of malaria each year. Of the four species of malarial parasites that infect humans, the two major species are Plasmodium falciparum and P. vivax.
The P. falciparum blood stage merozoites bind to and parasitize the erythrocytes using a variety of surface proteins (Cowman et al., FEBS Lett. 476:84-88 (2000); Baum et al., J. Biol. Chem. 281:5197-5208 (2006)), a major antigenic member of which is called Merozoite Surface Protein 1 (MSP1), a 195 kDa protein. MSP1 is present in all the erythrocyte-invasive species of Plasmodium, anchored to the merozoite surface by a glycosyl-phosphatidylinositol linkage. During early stages of the erythrocyte invasion process, soon after release from infected erythrocytes, the merozoite MSP1 protein undergoes proteolytic cleavage, producing a C-terminal cleavage product MSP1-42, which subsequently undergoes a second cleavage, producing an 11 kDa peptide MSP1-19, which remains attached to the parasite surface as it enters the erythrocyte. The formation of the cleavage product MSP1-19 is very important for successful invasion by the parasite since inhibition of its proteolytic formation or its neutralization by monoclonal antibodies prevents entry of the parasite to the erythrocytes (Blackman et al., J. Exptl., Med. 180:389-393 (1994)).
The MSP1-19 peptide is one of the most important malaria vaccine candidates available. MSP1-19-specific antibodies from malaria-resistant human sera react with the antigen and include a major erythrocyte-invasion inhibitory component (Holder & Riley, Parasitol. Today, 12: 173-174 (1996); O'Donnell et al., J. Expt. Med. 193: 1403-1412 (2001)). Serum from donors in malaria-endemic regions usually demonstrates strong antibody reactivity towards Pf MSP1-19. (Nwuba et al., Infect. Immun. 70: 5328-5331 (2002))
The monoclonal antibody (mAb) G17.12 was raised against recombinant Pf MSP1-19 and recognizes its epitope on the parasite surface, demonstrating that this region of the antigen is accessible on the native MSP1 polypeptide complex (Pizarro et al., J. Mol. Biol. 328:1091-1103 (2003)). Interestingly, erythrocyte invasion experiments in vitro showed that infection is not inhibited in the presence of G17.12, even at 200 μg/ml concentration and G17.12 does not inhibit in vitro secondary processing of MSP1. Id. The presence of antibodies that block the binding of invasion-inhibitory antibodies, thereby facilitating parasite survival, has also been demonstrated (Guevara Patino et al., J. Expt. Med. 186: 1689-1699 (1997)), and may be responsible for the failure of G17.12 mAb to inhibit erythrocyte invasion by M. falciparum.
Cerebral malaria, a rare but fatal infection restricted to P. falciparum invasion of brain capillaries because of the sequestration of parasitized erythrocytes, is often untreatable because most drugs cannot cross the blood-brain barrier to reach the brain capillaries. Adhesion of P. falciparum—infected erythrocytes to brain capillaries is mediated by the interaction of parasite ligands Pf Emp-1 family of proteins expressed on the surface of infected erythrocytes with ICAM-1 and CD36 expressed on the surface of capillary endothelium cells in cerebral vessels. (Smith et al., Proc. Natl. Acad. Sci. USA 97:1766-1771 (2000); Franke-Fayard et al., Proc. Natl. Acad. Sci. USA 102, 11468-11473 (2005))
Although a few drugs, such as chloroquine that targets the heme detoxification pathway, are used to treat malaria, there are increasing incidence of parasite resistance to drugs and mosquito vector resistance to insecticides. Chloroquine antagonizes heme polymerization mediated by parasite-induced HRPs (histidine-rich proteins), as heme monomers are highly toxic for malaria parasites. The polymerization of heme allows detoxification, which is reversed by chloroquine. Another drug, artemisinin, is effective against chloroquine-resistant P. falciparum in cerebral malaria. Artemisinin forms adducts with globin-bound heme in hemoglobin, which binds HRPs to prevent heme polymerization. There is an urgent need to find new drugs for this dreaded disease that is particularly prevalent in Africa and Asia. Present attempts at drug development are directed towards deciphering the complete parasite genome sequence, molecular modeling of the malaria parasite proteins and a search for novel drug targets.

Compositions and methods for treating malaria with cupredoxin and cytochrome

The Board of Trustees of the University of Illinois


A method and apparatus for converting an analog input into a final digital output. A first digital output is generated in response to receiving an analog input at a first converter. A second digital output is generated in response to receiving the analog input at a second converter. The first digital output is output to form the final digital output when an amplitude of the analog input is within a first portion of an overall range of amplitudes for the analog input and the second digital output is output to form the final digital output when the amplitude is within a second portion of the overall range of amplitudes.

1. Field
The present disclosure relates generally to analog and digital signals and, in particular, to analog-to-digital converters. Still more particularly, the present disclosure relates to a method and apparatus for converting an analog input into a digital output using a conversion device capable of multiple resolutions.
2. Background
An analog-to-digital converter (ADC) is a device that quantizes an analog signal. The analog signal may be a continuous input signal representing a measurable parameter such as voltage or current. Quantization is the process of converting a continuous range of values into a finite range of discrete values. In this manner, quantizing an analog signal means creating a series of discrete digital values that represent the analog signal. These discrete digital values may be represented in, for example, binary format.
The resolution of an analog-to-digital converter is determined by the number of discrete values that can be produced over a range of analog values. In some cases, the resolution of an analog-to-digital converter may be expressed in bits and referred to as bit resolution. An analog-to-digital converter with an N-bit resolution may be able to produce 2N discrete values.
In other cases, the resolution of the analog-to-digital converter may be defined based on the type of measurements received as input. For example, the resolution of an analog-to-digital converter that receives an analog voltage signal as input may be the overall voltage measurement range for the analog-to-digital converter divided by the number of discrete values possible. In particular, the voltage resolution of an analog-to-digital converter may be determined by the least significant bit (LSB) voltage, which is the minimum change in voltage needed to cause a change in the digital output. As the least significant bit voltage decreases, the resolution increases.
In some cases, one portion of a range of measurements may be of more interest than a rest of the range of measurements. For example, a resistance temperature sensor may contain a resistor that changes resistance as a temperature of the resistor changes. As the resistance changes, the voltage signal that is output from the resistance temperature sensor also changes.
However, the change in voltage based on a one degree change in temperature may be smaller at colder temperatures, such as the cryogenic range, as compared to hotter temperatures. Consequently, the discrete values generated by an analog-to-digital converter receiving the voltage signal from the resistance temperature sensor may have a higher degree of error with respect to the colder temperatures as compared to the hotter temperatures.
Some currently available analog-to-digital converters may be unable to improve this error without increasing the bit resolution of the analog-to-digital converter. Increasing the number of bits may increase the amount of digital data that needs to be processed, stored, or both more than desired. Therefore, it would be desirable to have a method and apparatus that take into account at least some of the issues discussed above, as well as other possible issues.

Method and apparatus for non-uniform analog-to-digital conversion


A latching mechanism for a sub-sea control module is constructed with energy storing means such that a latch is cooked into an unlatched condition by the action of lifting the sub-sea control module to store energy in the energy storing means and is moved into a latched condition by the action of lowering the sub-sea control module into abutment with the mounting base.

For an underwater oil well to become operational it is necessary to install at the well-head an assembly known as a "Christmas Tree" which combines equipment for monitoring and controlling the output flow from the well.
The Christmas Tree fitted to the well-head comprises, in particular, a large number of hydraulically actuated valves, the remote control of which is effected from a general operating station on the surface.
In particular, this operating station can be located on an oil rig which is connected to one or more underwater stations each by an umbilical cord providing the means for conveying electrical and/or hydraulic energy and the transmission of electrical or optical signals for controlling the oil extraction. The operating station may also be shore based.
Each underwater station may comprise one or more well-heads.
The operating station on the surface thus enables the remote control of a very large number of valves.
Each of the hydraulically actuated valves is of the type comprising a hydraulic valve actuator which is connected to a supply of pressurized fluid via a control unit, comprising a control valve for the flow of pressurized fluid and means for connecting the unit to the supply of pressurized fluid and to the network for transmission of control signals of the valve.
According to a known arrangement, the control of the operation of the assembly of hydraulically actuated valves of a Christmas Tree of one well-head is effected by a control unit secured to the well-head and which is connected to the hydraulic valve actuators by flexible pipes.
This control unit, which is known as a Sub-sea Control Module (SCM) or "POD", is a heavy and expensive apparatus which is specific to the configuration of a well-head.
Typically, such a unit is lowered onto the sub-sea installation using special Remotely Operated Vehicles (ROVs) and Remotely Operated Tooling (ROTs) from floating work barges or service vessels using soft landing guide wires and latching pins located on the sub-sea well installation. The weight of the control unit is typically 1.5 tons or more and requires substantial framing and counterweights on the installation to balance the loads on the well-head Christmas Tree.
The present invention is concerned with the mechanism used to releasably latch the so-called sub-sea control module, or pod to the Christmas Tree.
In a known arrangement this latching mechanism comprises a screw driven bolt-like member having a star-shaped lower end adapted to engage in a complementary shaped aperture in a pod mounting base carried by the Christmas Tree. Such a device is known as a retlock, is relatively expensive to manufacture and requires motive power to drive the rotatable screw in order to move it into and out of the locking or latching condition.
The present invention is concerned with simplifying and reducing the cost of this type of latching mechanism and making it more rapidly acting.


A liquid crystal display according to an embodiment of the invention includes a substrate, a plurality of gate lines formed on the substrate, a plurality of data lines formed on the substrate to intersect the gate lines, and a plurality of pixel electrodes formed on the substrate. In the liquid crystal display, the pixel electrode includes a first main side substantially parallel with the gate line, a second main side substantially parallel with the data line, a first oblique side making a first oblique angle with respect to the first and second main sides, and a second oblique side making a second oblique angle with respect to the first and second main sides. The first oblique angle and the second oblique angle are different from each other.

A liquid crystal display is one of numerous types of flat panel displays that are being widely used. A typical liquid crystal display includes two display panels having corresponding electric field generating electrodes such as pixel electrodes and a common electrode formed thereon, and a liquid crystal layer interposed therebetween. When a voltage is applied to the electric field generating electrodes, an electric field is generated in the liquid crystal layer. The electric field determines the alignment of liquid crystal molecules of the liquid crystal layer, which controls the polarization of incident light. In this way, images are displayed.
The liquid crystal display further includes switching elements connected to individual pixel electrodes, and a plurality of signal lines such as gate lines and data lines for controlling the switching elements to apply a voltage to the pixel electrodes.
Among the various types of liquid crystal displays, a longitudinally-aligned-mode liquid crystal display is popular because it has a high contrast ratio and a wide reference viewing angle. In the longitudinally-aligned-mode liquid crystal display, when no electric field is applied, the major axis of each liquid crystal molecule is aligned to be longitudinal to a display panel.
Specific methods of realizing a wide viewing angle in the longitudinally-aligned-mode liquid crystal display include a method of forming cutouts in electric field generating electrodes and a method of forming protrusions on electric field generating electrodes. The cutouts and the protrusions determine the tilt directions of the liquid crystal molecules. Therefore, when the cutouts and the protrusions are suitably disposed so as to disperse the tilt directions of the liquid crystal molecules in various directions, it is possible to achieve the wide reference viewing angle.
In such a liquid crystal display, in order to raise luminance, the strength of the electric field formed in the liquid crystal layer may be increased. However, when an applied voltage to the pixel electrodes is raised in order to increase the strength of the electric field, the electric field formed between the pixel electrodes and the data lines for transmitting the voltage to be applied to the pixel electrodes also becomes stronger. This strong electric field disorders the alignment of the liquid crystal molecules positioned around the pixel electrode edges, increasing the response time of the liquid crystal.
Meanwhile, since it is difficult for light to pass through portions where the protrusions or the cutouts exist, the more protrusions or cutouts, the lower the aperture ratio. However, when the separation between the protrusions or the cutouts increases in order to raise the aperture ratio, the effects of the protrusions or the cutouts are relatively decreased and disturbance in the electric field due to the data lines becomes severe, thus increasing the response time.
In a rectangular liquid crystal display in which pixel electrodes are parallel with gate lines and data lines, an electric field generated between neighboring pixel electrodes disorders the alignment of liquid crystal molecules. As a result, a texture is generated. Accordingly, the transmittance is decreased and a residual image remains on a screen.
In order to reduce such texture, a cutout of a common electrode may overlap a side of a pixel electrode. However, the aperture ratio may be lowered, which may have the undesirable effect of reducing transmittance.


A cell, in particular a button cell, and to a method for manufacturing such a cell, the method includes providing a first part and a second part intended to respectively form the lid and the cup of the housing, the first part including an edge area with a zone inclined or perpendicular with respect to a center axis of the housing; applying a layer of adhesive on the edge area of the first part; then inserting the first part into an open end of the second part, the layer of adhesive on the edge area being finally turned towards the open end of the second part; closing the housing by bending an upper portion of the side wall on the zone of the edge area provided with the layer of adhesive, and curing the layer of adhesive to form an adhesive joint sealing the housing.

Button cells usually have a housing with a cup and a lid respectively forming the positive and the negative poles. Generally, a preformed elastomer seal arranged between the cup and the lid ensures the galvanic isolation between both parts whilst the liquid-tight closure of the cell is carried out by crimping the edge of the cell cup over the cell lid to compress the seal. This current design of the housing 2 is represented in FIG. 1 with the cup 3, the lid 4 and the seal 5. This construction of the housing with an elastomer seal takes up a lot of space, which reduces the volume available for the active material. For this standard construction, the ratio between the internal volume of the housing available for the active material and the external volume of the housing only amounts to ˜70%.
In order to increase the internal volume and thus the capacity of the cell, a solution is to replace the elastomer seal with an adhesive joint. This solution is promising but needs further improvements in particular in terms of adhesion between the assembled parts. Several factors can adversely affect the adhesion of the joint and hence the tightness of the cell. More specifically, in use, the joint can be subject to a stress in case of overpressure inside the cell. Depending on the position and the orientation of the joint, the stress can have a positive or a negative impact on the adhesion of the joint. So, if the joint is not adequately arranged, it will be subject to a tensile and/or a shear stress impairing its adhesion.
Other factors during the manufacture of the cell can also affect the adhesion of the joint. For example, the presence of liquid active material such as the electrolyte inside the housing can contaminate the surfaces to be assembled and thereby compromise the adhesion of the joint on these surfaces.

Cell and method for manufacturing such a cell


A gaze tracking model is adapted to predict a gaze ray using an image of the eye. The model is trained using training data which comprises a first image of an eye, reference gaze data indicating a gaze point towards which the eye was gazing when the first image was captured, and images of an eye captured by first and second cameras at a point in time. The training comprises forming a distance between the gaze point and a gaze ray predicted by the model using the first image, forming a consistency measure based on a gaze ray predicted by the model using the image captured by the first camera and a gaze ray predicted by the model using the image captured by the second camera, forming an objective function based on at least the formed distance and the consistency measure, and training the model using the objective function.

Different techniques have been developed for monitoring in which direction (or at which point on a display) a user is looking. This is often referred to as gaze tracking or eye tracking. Such techniques often involve detection of certain features in images of the eye, and a gaze direction or gaze point is then computed based on positions of these detected features. An example of such a gaze tracking technique is pupil center corneal reflection (PCCR). PCCR-based gaze tracking employs the position of the pupil center and the position of glints (reflections of illuminators at the cornea) to compute a gaze direction of the eye or a gaze point at a display.
As an alternative (or complement) to conventional techniques such as PCCR-based gaze tracking, machine learning may be employed to train an algorithm to perform gaze tracking. More specifically, the machine learning employs training data in the form of images of the eye and associated known gaze points to train the algorithm, so that the trained algorithm can perform gaze tracking in real time based on images of the eye. Plenty of training data is typically needed for such machine learning to work properly. The training data may take quite some time and/or resources to collect. In many cases, certain requirements may be put on the training data. The training data should for example preferably reflect all those types of cases/scenarios that the gaze tracking algorithm is supposed to be able to handle. If only certain types of cases/scenarios are represented in the training data (for example only small gaze angles, or only well-illuminated images), then the gaze tracking algorithm may perform well for such cases/scenarios, but may not perform that well for other cases/scenarios not dealt with during the training phase.
It would be desirable to provide new ways to address one or more of the abovementioned issues.


A universal joint has a ball retainer having a retainer body made of a structurally rigid material and a relatively soft elastomeric seal bonded to the body at an axial end in a recess so as to wipe lubricant back on the ball that is retained by the body. Cavity plates in a housing of the universal joint partially close off lubricant reservoir recesses to retain lubricant in the reservoirs and act as wear plates that stabilize the centering plate orbiting in the recess. Seals can be provided between the retainer and the ball socket, between the centering plate and the wear plates and between the wear plates and the cavity plates. Passageways connect the center cavity of the housing with the ball sockets through an orifice.

It is well known that universal joints, and in particular high angle double cardan universal joints, need lubrication over a prolonged period of high speed operation, without throwing the lubrication out due to centrifugal and other forces on the universal joint. This is discussed in U.S. Pat. Nos. 4,781,662 and 5,688,065, the disclosures of which are hereby incorporated by reference for their disclosures of universal joints, and in particular lubrication of universal joints.
Although universal joints are well known in the art, a problem persists in retaining lubricant in the joint for as long as possible. Typically, the joint is lubricated when new. After being placed in service, the joint may never be lubricated again. The joint is subjected to centrifugal forces in service, which tend to cause the lubricant to be thrown radially outward. The invention described herein is accordingly directed to retaining lubricant in the joint during operation.