The present invention provides a new approach for cancer treatment by utilizing gene therapy combined with radiation therapy to enhance cytotoxicity in malignant cells. Specifically, the present invention demonstrates that molecular chemotherapy with the cytosine deaminase gene and 5-fluorocytosine is an effective radiosensitizing strategy which may lead to substantial improvement in tumor control, with less normal tissue toxicity than conventional systemic administration of 5-fluorouracil, that would translate into improved cure rates and better survival. Also provided is a noninvasive method for continuous in vivo monitoring of 5-fluorouracil production via magnetic resonance spectroscopy.

The present invention relates generally to the fields of molecular biology, radiation oncology and cancer therapy. More specifically, the present invention relates to the finding that a combination of molecular chemotherapy and radiation therapy enhances therapeutic effects against cancer.
Clinical applications of cancer gene therapy have had limited success due to a variety of factors, including ineffective therapeutic gene delivery in situ. The physiologic milieu of the target tumor may have deleterious effects on the delivery of therapeutic genes. This limitation may be disease specific, and variable depending on the specific tumor type and tumor location. Most clinical gene therapy trials thus far have utilized compartmental models of malignant disease (1, 2). In this regard, thoracic malignancies and intra-abdominal carcinomatosis represent common body compartmentalized diseases that have been explored in an experimental therapeutic context. Attempts to address the issue of achieving viral vector delivery to cancer cells in the face of a physiologic infection medium of pleural fluid or abdominal ascites have been examined (3, 4). Yang et al. demonstrated retroviral transduction of pancreatic cancer cells in the presence of human ascites, which was similar to the results obtained in culture medium (3). Batra et al. reported significant inhibition of retroviral transduction of mesothelioma cells in the presence of malignant pleural fluid, specifically the chondroitin sulfate proteoglycan fraction (4).
Radiotherapy combined with the radiosensitizing chemotherapeutic drug 5-fluorouracil (5-FU) has been studied as a therapeutic modality in many human tumor types (5). Systemic toxicity limits the amount of 5-FU that can be administered for many clinical anti-cancer applications (6, 7). Radiation therapy and gene therapy have the potential to be combined to enhance effectiveness of cancer therapy without enhancing dose limiting toxicity. To this end, reports have investigated this interaction (8). These include: TNFxcex1 under the control of a radiation inducible promoter (9, 10), conversion of prodrugs to toxic metabolites that are also radiosensitizers (11-15), p53 mediated radiosensitization (16, 17) and the genetic induction of membrane receptors that can b e targeted with radiolabeled peptides (18-21).
With respect to enzymatic conversion of nontoxic prodrugs into radiation sensitizing agents, the genes for bacterial and yeast cytosine deaminase (CD) have been cloned and studied (22, 23, 40). Cytosine deaminase converts a nontoxic prodrug 5-fluorocytosine (5-FC) into 5-FU. The cytosine deaminase gene has been used in gene therapy strategies to mediate intracellular conversion of 5-FC to 5-FU, and has been shown to be effective in animal tumor models of human colon carcinoma (24). Human colon cancer cells that have been stably transduced to express the cytosine deaminase gene have been shown to be radiosensitized by the addition of 5-FC in vitro and in vivo (13). Adenoviral vectors have been used to achieve efficient gene delivery in a variety of tissues in vitro and in vivo. Adenoviral vectors encoding the cytosine deaminase gene have been described (25, 26).
Presently available assays for determining intratumoral 5-FU concentration are problematic. They require the removal of a tumor, the homogenization of that tumor and the collection of the cellular lysate in order to directly measure 5-FU concentration, usually by high-pressure liquid chromatography. No noninvasive method of detection existed, which could allow for continuous in vivo monitoring of 5-FU production.
In the context of multiple administrations of adenoviral vectors, the host immunologic response, with generation of neutralizing anti-adenovirus antibodies and cytotoxic T cells, is thought to limit the potential effectiveness of secondary administration of adenoviral vectors. A means to overcome this problem may be to improve the effectiveness of infection of the initial viral challenge, i.e., to enhance the transduction efficiency of the adenoviral vector for the target cells at the initial adenoviral administration. This goal may be achieved by utilizing a ligand to a cellular receptor overexpressed in the target carcinoma cells to redirect adenovirus vector binding.
The prior art is deficient in the lack of effective means of treating of human cancers by chemotherapy combined with radiation therapy to produce enhanced therapeutic effects against cancer and reduced normal tissue toxicity. In addition, the prior art is deficient in the lack of effective means of redirecting adenovirus vector binding via a cellular receptor to improve the effectiveness of gene therapy. Furthermore, the prior art is deficient in the lack of a noninvasive method for continuously monitoring therapeutic transgene expression in tumors therefore improving the gene therapy. The present invention fulfills this long-standing need and desire in the art.
The present invention is directed to a method of transfecting established tumors in vivo with an adenovirus encoding the cytosine deaminase gene, administration of systemic 5-FC, and radiation therapy, (e.g., external beam or brachytherapy) of the tumor. This method results in tumor regression and prolonged tumor growth inhibition compared to control treatments with molecular chemotherapy or radiation therapy alone. Also disclosed is an adenoviral-conjugate mechanism to circumvent current limitations of cancer gene therapy to solid gastrointestinal malignancies.
Specifically, the present invention utilizes an adenoviral vector under the control of a cytomegalovirus promoter (AdCMVCD) encoding cytosine deaminase in combination with 5-FC and single fraction radiotherapy to demonstrate enhanced cytotoxicity to WiDr human colon carcinoma cells in vitro. The present invention also demonstrates such gene therapy/prodrug treatment strategy employing a fractionated radiation dosing schema in animal models of WiDr human colon carcinoma and SK-ChA-1 human cholangiocarcinoma. A prolonged WiDr tumor regrowth delay was obtained with AdCMVCD infection in combination with systemic delivery of 5-FC and fractionated external beam radiation therapy compared to control animals treated without radiation, without 5-FC, or without AdCMVCD. The present invention further discloses redirection of adenovirus vector (AdCMVCD) binding via a ligand to a cellular receptor, e.g., the basic fibroblast growth factor (FGF2) receptor, to improve the effectiveness of gene therapy in combination with 5-FC treatment and radiation therapy.
Clinical applications for cancer gene therapy are limited by the inability to genetically modify a majority of tumor cells to achieve a therapeutic effect. In this regard, the enzyme/prodrug strategy consisting of cytosine deaminase/5-fluorocytosine (CD/5-FC) relies on diffusion of the cytotoxic enzymatic product 5-FU to kill non-transduced tumor cells. Methods to increase solid tumor transduction in situ may augment therapeutic gene expression and response to therapy. To this end, gene delivery was improved via vector binding to molecules expressed on the cells of tumors. Fibroblast growth factor (FGF) receptors are overexpressed in a majority of pancreatic carcinomas, but poorly characterized in cholangiocarcinoma. Targeted adenovirus via basic fibroblast growth factor (FGF2) to the fibroblast growth factor receptor was used as a vehicle for the delivery of cytosine deaminase to hepatobiliary tumor cells for combination molecular chemotherapy and radiation therapy studies.
FGF2 redirected adenoviral delivery of firefly luciferase gene (AdCMVLuc) expression was evaluated in vitro. Transduction efficiencies using adenoviral delivered E. coli xcex2-galactosidase gene (AdCMVLacZ) expression also was determined. The methodology to redirect adenoviral gene delivery employed the Fab fragment of a neutralizing anti-adenoviral knob monoclonal antibody which ablates native adenoviral tropism, conjugated to FGF2 ligand which provides for FGF receptor binding. An adenoviral vector encoding the cytosine deaminase gene (AdCMVCD), in combination with 5-FC and the Fab-FGF2 conjugate, was used to evaluate differential cytosine deaminase protein expression by Western blotting of transfected cell lines and enzymatic activity by increased conversion of 3H-5-FC into 3H-5-FU. Proliferation assays were performed to correlate differential production of 5-FU with increased cytotoxicity in selected pancreatic and cholangiocarcinoma cell lines. In vivo studies utilizing AdCMVCD, the Fabxe2x80x2-FGF2 conjugate, 5-FC administration, and a single 5 Gy dose of external beam radiation to the tumor in nude mice were performed to evaluate the anti-tumor efficacy of AdCMVCD+Fabxe2x80x2-FGF2, compared to AdCMVCD alone, in established subcutaneous BXPC-3 pancreatic tumors.
In target cells, FGF2 retargeted AdCMVLuc resulted in enhanced (10-100-fold) levels of firefly luciferase expression relative to AdCMVLuc infection alone. X-gal staining for xcex2-galactosidase expression revealed an enhanced transduction frequency mediated by Fab-FGF2 redirected AdCMVLacZ compared to AdCMVLacZ infection alone. Fab-FGF2 redirection of AdCMVCD resulted in increased cellular expression of cytosine deaminase and production of 5-FU, and enhanced cellular cytotoxicity at low viral multiplicities of infection, compared to the levels obtained with AdCMVCD alone. In BXPC-3 tumor-bearing animals treated with AdCMVCD+Fabxe2x80x2-FGF2, 5-FC, and radiotherapy, the time to tumor size doubling was extended compared to AdCMVCD, 5-FC, and radiotherapy alone.
These results indicate that native adenoviral tropism can be redirected using ligands to cell surface receptors. In addition, transduction efficiencies and expression of novel genes introduced via this heterologous pathway are significantly enhanced compared to native adenovirus transduction alone. These findings suggest improved gene expression may be achieved via this adenoviral-conjugate mechanism to circumvent current limitations of cancer gene therapy to solid gastrointestinal malignancies.
The present invention is further directed to a noninvasive method for monitoring the continuous conversion of 5-fluorocytosine to 5-fluorouracil via magnetic resonance spectroscopy (MRS). Magnetic resonance spectroscopy allows for monitoring this prodrug activation therapy through the following: the identification of tumor and normal tissue sites of production or accumulation of 5-fluorouracil, the discrimination of both 5-fluorocytosine clearance/5-fluorouracil production, the determination of the residence time of 5-fluorouracil, the production of metabolites of the active drug, along with the determination of the elimination kinetics of 5-fluorouracil from tumor and normal organs. The information that magnetic resonance spectroscopy can provide about the pharmacokinetics of these agents can help develop procedures to maximize the effectiveness of this therapy with the potential to maximize tumor regression.
In one embodiment of the present invention, there is provided a method of treating an individual having a solid tumor, comprising the steps of treating the individual with an adenovirus encoding a cytosine deaminase gene; administering 5-fluorocytosine to the individual; and treating the individual with radiation therapy.
In another embodiment of the present invention, there is provided a method of treating an individual having a cancer, comprising the steps of combining a ligand that binds to a tumor cellular receptor and an adenoviral vector encoding a cytosine deaminase gene to form a complex; treating the individual with the complex; administering 5-fluorocytosine to the individual; and treating the individual with external beam irradiation.
In still another embodiment of the present invention, there is provided a method of monitoring continuous conversion of 5-fluorocytosine to 5-fluorouracil in a tumor, wherein the tumor is treated with multiple doses of 5-fluorocytosine and multiple doses of adenovirus encoding a cytosine deaminase gene, comprising the steps of placing the treated tumor in a magnet; and evaluating the presence of 5-fluorocytosine and 5-fluorouracil by magnetic resonance spectroscopy over a course of time, wherein less amount of 5-fluorocytosine and more amount of 5-fluorouracil indicates increased conversion of 5-fluorocytosine to 5-fluorouracil. Preferably, the tumor is further treated with radiation.
In still yet another embodiment of the present invention, there is provided a method of monitoring continuous conversion of 5-fluorocytosine to 5-fluorouracil in a tumor, wherein the tumor is treated with multiple doses of 5-fluorocytosine and multiple doses of cytosine deaminase gene encoding adenovirus targeted by a ligand to a tumor cellular receptor, comprising the steps of placing the treated tumor in a magnet; and evaluating the presence of 5-fluorocytosine and 5-fluorouracil by magnetic resonance spectroscopy over a course of time, wherein less amount of 5-fluorocytosine and more amount of 5-fluorouracil indicates increased conversion of 5-fluorocytosine to 5-fluorouracil. Preferably, the tumor is further treated with radiation.
Other and further aspects, features, and advantages of the present invention will be apparent from the following description of the presently preferred embodiments of the invention given for the purpose of disclosure.

Molecular chemotherapy enhancement of radiotherapy


An air purifier which includes an air intake with electrically charged plates for ionizing particles of smoke, dust, and impurities entrained in the air, a fan or other air-moving device to move the polluted air through the intake and over the ionizing plates and to discharge the air against a collector which is spaced from the charged plates and is electrically grounded and upon which the ionized pollutants will deposit, and an air outlet through which the purified air can be discharged, leaving the pollutants behind on the collector element.

Electrostatic precipators in general are not new, nor is the principle for clarifying air by the use of electrostatic precipitators.
Such prior precipitators or air purifiers have, however, in the past, been fairly large, expensive, and cumbersome, not portable, and usually constructed for permanent installation in a built-in air conduit. This rendered the device relatively limited in its operation or required the purchase of multiple units for use in different areas of the air ducts.


A computer system, a computer-implemented method, and a computer program product determine an operating state of at least one system component in a fluid-carrying, cascading production system. The computer system includes an interface configured to receive a first and a second pressure pulse signal. The first and second pressure pulse signals characterize, respectively, a pressure pulse before and after the pressure pulse passes through the production system. A memory unit stores a plurality of system component-specific transmission functions which describe an operating-state-dependent pulse response of the at least one system component to the pressure pulse. An evaluation unit determines a real transmission function from the first and second pressure pulses, and compares the real transmission function of the production system with the stored plurality of transmission functions for the system components.

1. Field of the Invention
The invention relates to electronic data processing in general and specifically to a computer system, a computer program product and a computer-implemented method for monitoring components of a fluid-carrying, cascading production system.
2. Description of Related Art
Sensors for monitoring specific parameters are used after each production step and cleaning step in fluid-carrying, cascading production systems, such as water-processing systems for dialysis. For each sensor-monitored component of the production system, these sensors supply measured data that can be used to determine the operating status of the respective components. One example of such a specific parameter is the pressure difference between the input pressure of a specific component and the output pressure of that specific component. For example, when the sand filter is filled with cleaned sand, this pressure difference is different from that when the sand filter is contaminated with particles washed into it. An inference about the operating status of the respective component can thus be drawn from the specific parameter (e.g., pressure difference). In the case of monitoring of the specific parameters for the individual components of the cascading production system, it is possible to monitor the functionality of the production system remotely and optionally to plan and prepare for service or repair jobs. Such complex monitoring systems generally use two sensors per component, where the first sensor measures the input value of the specific parameter and the second sensor measures the output value of the parameter for the respective component. A great many sensors are thus required for cascading production systems, thus resulting in a great system complexity and also a large volume of measured data.

Monitoring system and method for fluid-carrying production system


A measurement system wherein an acoustic distance measuring transducer is moved over a flat reference surface, measuring the distance from the transducer to a calibration surface and then to a test surface. The transducer is moved along a path within a ring while the top edge of the ring is used to support the calibration surface and then the test surface. The ring is mounted for rotation so that the transducer can scan a supported surface at a locus of points. Scans across the calibration surface and then the test surface are compared to measure deviations of the test surface from calibration surface.

a. Field of the Invention
The invention relates to non-contacting measurement of surfaces, and in particular to a scanning acoustic measurement system for testing flatness of wafer surfaces and the like.
b. Prior Art
Previously, non-contacting measurements of flatness for wafers and similar articles have been made optically and capacitively. With respect to optical devices, reflective and interferometer devices are known. One reflective type employs a beam which is directed onto a test surface from a position perpendicular to the surface. Small deviations in flatness cause the beam to be reflected at an angle which is not perpendicular to the surface. By measuring the angle of reflection plus the duration and extent of the reflective deflection as the beam is moved along a straight line, the deviation from flatness may be computed.
In the interferometer instrument, fringes are created by means of splitting a beam of monochromatic light from a single source into two components of which one follows a path which includes a reflection on a test surface and the other follows a path which includes a reflection on a reference surface. The path length difference causes an optical fringe pattern to appear. Any deviation in flatness causes the number of fringes to change. By counting fringes, the deviation from flatness may be measured. A problem with optical methods is that specularly reflective surfaces are required.
Another apparatus for measuring flatness of a test surface is a capacitive gauge which consists of three spaced apart reference sensors, together with an array of similar sensors. The three reference sensors are used to capacitively determine the distance to a test surface. The plane defined by those three distances is a reference plane. The array of other sensors then measures the distance to the test surface and any deviations from the reference plane indicate deviations in flatness. A problem with capacitive measurement devices is that the test surface must be at least partially conductive.
In prior U.S. Pat. No. 4,175,441, K. Urbanek, G. Kren and W. Wheeler disclose an acoustic gauge for measuring distances to planar members and the thickness of flat sheets and the like.
While prior art devices make generally accurate measurements of flatness, in most instances the time for obtaining an accurate multi-point flatness appraisal of a test object is limited. This is because most devices measure either a single point at a time or a modest number of points simultaneously, except for the laser reflective apparatus. In the case of that instrument, the laser beam scans one line across the surface, but time must be taken to determine the extent of a slope, as indicated by a variance in the angle of reflection of the beam. In the case of the interference apparatus, an entire surface may be measured at a time, but interpreting the data by counting fringes and measuring spacing between fringes is tedious.
An object of the invention is to devise a non-contacting flatness gauge wherein a flatness profile based on a large number of sample points may be obtained in a relatively brief time interval.

Acoustic method and apparatus for measuring surfaces of wafers and similar articles


An automatic focusing camera having a ranging device, which selects the nearest subject for focusing, including a manually operative member for defeating the ranging device and automatically signalling the lens drive arrangement to move the lens to a preselected focal position when focusing on other than the near subject is desired; as, for example, when focusing through a window with a sonic ranging camera.

1. Field of the Invention
This invention relates generally to the field of focusing apparatus and, more particularly, selectively actuatable apparatus for automatically causing a variable focus system to have a lens element positioned at a preselected focal position, and for suppressing firing of a source of artificial illumination.
2. Description of the Prior Art
Ultrasonic ranging systems are well-known for use in controlling operations in photographic apparatus. Essentially, ultrasonic energy is transmitted to a subject and the reflected signals from the subject are received by the system. Characteristics of the transmitted and received signals are compared for purposes of deriving a ranging signal indicative of the subject distance. Such signal may be used for purposes of driving an element of a variable objective lens assembly of the photographic apparatus to a position corresponding to the subject distance, whereby the subject will be in focus at a focal plane in the photographic apparatus.
An example of such a system is disclosed in U.S. Pat. No. 4,199,246, entitled "Ultrasonic Ranging System for a Camera", by J. Muggli, assigned in common herewith. This patent describes use of an ultrasonic transducer for transmitting a relatively short burst of ultrasonic frequencies, and a synchronized receiver for processing an echo signal produced by the transducer upon receipt of the echo after a predetermined time interval. This receiver produces a ranging signal having a characteristic related to the distance of the nearest subject being photographed. In response to the ranging signal, a lens element of a variable focus lens system is moved to the appropriate focal position corresponding to the subject range.
Despite several significant advantages of ultrasonic ranging systems and the widespread commercial acceptance thereof, certain photographic conditions exist wherein there are potential for problems. For example, focusing difficulties arise when the object to be focused is not the nearest object in the scene, for example, when it is situated beyond an acoustically opaque, but visually transparent material, such as a window. This is because the rangefinder will range on the window rather than the object beyond the window. Obviously, the desired subject will be misfocused at the focal plane.
Towards the end of correcting such problems efforts have been undertaken to provide these types of automatic focusing systems with an override system enabling independent manual control of the focusing system. Examples of such efforts are described in U.S. Pat. Nos. 4,149,792, 4,153,903 and 4,160,590, all commonly assigned herewith.
Thus, the foregoing patents disclose a manual control system for focusing in addition to an automatic focusing system. In such systems a manual focus wheel is operable for achieving the manual focusing when the automatic rangefinder is disabled. Disabling of the automatic focus control system may be effected by rendering the automatic focus control insensitive to the output signal from the rangefinder, by disabling the rangefinder itself or by any combination thereof.
In U.S. Pat. No., 4,149,792, a spring force biasing means is operative in the manual focusing mode for urging the lens from a position outside its normal focusing range to a position inside the normal focusing range to provide correct operation when the camera is later returned to the automatic mode. This latter positioning is therefore done to facilitate the automatic focusing mode and, consequently, does not directly overcome the above-noted problem associated with misfocusing of automatic rangefinders. Commonly assigned U.S. Patent No. 4,160,590 discloses a lens movement limiting stop that limits manual movement of a lens to within its normal focusing range. This patent, like other prior art, requires focusing control by the operator when faced with the problem of focusing upon objects located behind acoustically opaque and visually transparent material.

Apparatus for automatically selecting a preselected focal position for a lens system


A stereoscopic display device and a stereoscopic display method are provided. The stereoscopic display device comprises: a display unit configured for displaying an image and emitting emergent light; and at least two active scattering panels arranged at a light-emitting side of the display unit and in different planes. The active scattering panels are configured for performing a scattering process to the emergent light, so that the emergent light, after being scattered by the different active scattering panels, forms a stereoscopic picture. By providing a plurality of active scattering panels to achieve stereoscopic display without the provision of a plurality of display panels, it is possible to avoid the problem that backlight emitted by a backlight source is subjected to a serious reduction in transmittance after passing through a plurality of layers of display panels, thereby improving display brightness of a stereoscopic display device.

For the present, conventional 3D (three dimensional) stereoscopic display techniques may be distinguished into two main categories, i.e., the spectacle type 3D and the naked-eye type 3D. However, both the naked-eye type 3D and the spectacle type 3D display techniques utilize the parallax of the human eyes to achieve a 3D presentation, and therefore, are also called pseudo-3D. True-3D techniques mainly comprise the holographic imaging technology, the integrated imaging technology and the spatial 3D technology etc, the holographic imaging technology and the integrated imaging technology are relatively complicated, and the spatial 3D technology has a relatively simple manner in its implementation.
FIG. 1 is a schematic structural view illustrating a current stereoscopic display device. As illustrated in FIG. 1, the stereoscopic display device comprises a backlight source 1 and a plurality of layers of display panels 2 arranged in front of the backlight source 1, and the plurality of layers of display panels is arranged to be overlapped with each other. Because the display panels 2 are arranged in different planes, the pictures displayed by the different display panels 2 are located in different planes, so as to form a stereoscopic picture.
A true-3D display technique as described above is achieved by using the plurality of layers of display panels. However, due to the fact that the plurality of layers of display panels are arranged to be overlapped with each other, the light emitted by the backlight source is subjected to a severe reduction in transmittance after passing through the plurality of layers of display panels, and this causes the stereoscopic display device to have decreased display brightness.

Stereoscopic display device comprising at least two active scattering panels arranged in different planes each having a scattering function and a transmission function and stereoscopic display method


The present invention relates to a method for enriching nucleic acids with a length of not more than 300 nucleotides. The invention also relates to a kit for enriching nucleic acids with a length of not more than 300 nucleotides, to the use of such a kit, to the use of an anion exchange matrix and to a method for treating a disease.

1. Field of the Invention
The present invention relates to a method for enriching nucleic acids with a length of less than 300 nucleotides, to a kit for enriching nucleic acids with a length of less than 300 nucleotides, to the use of said kit, to the use of an anion exchange matrix and to a method for treating a disease.
2. Description of Related Art
Since small ribonucleic acids (RNAs) were found, a few years ago, to carry out an essential regulatory function in gene expression, scientific studies have increasingly concentrated on small RNAs shorter than 300 nucleotides, in particular shorter than 100 nucleotides. More specifically, many researchers have become interested in micro RNAs (miRNAs). miRNAs are an evolutionarily conserved class of small noncoding RNAs of about 22 nucleotides in length, whose complex role during regulation of gene expression is becoming more and more obvious. miRNAs have been found in any eukaryotic organism studied and in nearly every tissue, inter alia in fungi, plants, insects and mammals. Aside from miRNAs, other small RNAs have also been found which likewise play a fundamental role in cellular functions. These include, for example, small interfering RNAs (siRNAs), small nuclear RNAs (snRNAs) and small nucleolar RNAs (snoRNAs).
These short RNAs which are shorter than 300 nucleotides must be as pure as possible and purified with high yield from the biological systems to be studied in order for it to be possible for their cellular role to be investigated. There is therefore a great need to provide a method which enables such short RNAs to be purified or isolated from complex biological systems, in particular from cell lysates.
Generally, in order to isolate nucleic acids from biological samples, they must be separated from the remaining cellular components such as proteins, sugars, lipids and other components. The prior art has disclosed a plurality of methods of separating nucleic acids from very different starting materials, for example from cell cultures, from tissues of plant and animal origin and from body fluids. One method, for example, includes extracting the usually aqueous starting solutions with the aid of organic solvents such as phenol and chloroform (Chomczynski and Sacchi, 1987) followed by precipitation of the nucleic acids with the aid of alcohols such as ethanol or isopropanol from the aqueous phase (Sambrook, J., Fritsch, E. F. in T. Maniatis, CSH, "Molecular Cloning", 1989). Another method comprises immobilizing the nucleic acids to a solid phase, for example by means of silica adsorption technology. Disadvantageously, it is not, or only insufficiently, possible in all of these methods to isolate or at least purify relatively small nucleic acids.
To solve this problem, the prior art has disclosed methods for specifically enriching small RNA populations, which methods are based on silica membrane technology. In these methods, a specific, relatively small amount of alcohol is added to the cell lysate after the cells have been lysed, and at least part of the relatively long nucleic acids are bound to the silica membrane under chaotropic binding conditions. However, the amount of alcohol in the purification methods described in the prior art is too low to bind efficiently also small nucleic acids to the silica membrane, and these small RNAs are therefore present in the breakthrough. The alcohol concentration is then increased in the breakthrough and the latter is bound to a second silica membrane. After washing steps, the small RNA is eluted together with all other nucleic acids that have not been bound to the first column (see, for example, mirVana® kit from Ambion, Austin, USA or RNeasy® Lipid Tissue Mini Kit from QIAGEN, Hilden, Germany, to be used by means of the "user developed protocols").
The disadvantage of both the QIAGEN and the Ambion methods is that of two solid phases having to be used and it not being possible to obtain only the desired small RNA with the aid of a single binding step. Moreover, this method does not enable, for example, only miRNAs of about 22 nucleotides in size to be isolated, without isolating also tRNAs and other larger nucleic acids at the same time. Although it is possible to enrich the small RNA, in particular the miRNA, to a certain degree by this method under some circumstances, said small RNA is still contaminated with other nucleic acids, in particular with transfer RNA (tRNA).

Method for enriching short-chain nucleic acids


A body on frame vehicle includes a vehicle frame having a small offset impact load management system including upper and lower rear blocker structures and a reinforcement blocker structure for managing impact loads applied to the wheel and tire of the vehicle from being directed further toward the body. The reinforcement blocker structure includes a base member welded to openings in the frame side rail and extending angularly outwardly and located rearward and distal the wheel and tire and proximal a longitudinal cross frame member for transferring the small offset impact loads transferred by the wheel and tire in a cross vehicle direction.

It is generally known to provide a vehicle including a vehicle frame assembly of any known or appropriate type such as a unitary body or a body on frame assembly. It is also generally known to provide a vehicle including various structures for improving the performance of the vehicle during a variety of types of impacts to the vehicle. The performance of a vehicle and its various structures, assemblies and components from an impact may be assessed using a variety of crash tests and analytical methodologies.
A frontal crash having a relatively small amount of overlap may be designed to attempt to replicate what may happen when only a relatively small portion of the front corner of a vehicle collides with another object like a vehicle, tree, utility pole or the like. One known industry test is the small overlap rigid barrier (SORB) test. In the SORB test, a vehicle travels at 40 mph toward a 5-foot-tall rigid barrier and only the outer 25% of the vehicle width is impacted into the barrier. It is generally understood that most modern vehicles may be designed to have safety cages and other structures, assemblies and components for protecting the occupant compartment and built to help manage energy with controlled and limited deformation to the vehicle during a variety of impacts to the vehicle from most direction, including a head-on and overlap frontal crashes. The crush zones of the main body and frame structures are designed to manage the crash energy to reduce forces on the occupant compartment and its occupants. When a crash involves these structures, the occupant compartment may generally be protected from intrusion, and the airbags and safety restraints may perform to restrain and help protect vehicle occupants.
Small overlap frontal crashes primarily affect a vehicle's outer edges, which may not be directly protected by some of the primary crush-zone structures. In such a scenario, crash forces may go directly into the front wheel, suspension system and potentially the vehicle firewall and body including the passenger compartment. In a small overlap crash which does not engage the main structures of the vehicle it may be possible for the wheel to be forced rearward toward the passenger compartment of the body of the vehicle.
The wheel may be trapped between the rigid barrier and the body structure thus applying local loads that may surpass steel or aluminum structure material strengths. This is notably different since the understood prior art devices are designed and intended to prevent the wheel from moving, turning or twisting during a frontal impact and/or SORB testing. Without this invention, the wheel can get trapped between the rigid barrier and the body structure that applies local loads that may surpass steel or aluminum structure material strengths.
Even though such crush-zone and body (or body) on frame type structures have been known and may have some certain advantages, there remains a continuing and significant need to provide improved impact or crush performance structures having a lower cost structure and having optimized structural efficiencies. In body-on-frame vehicle architectures, particularly as the vehicle weight increases (such as with larger trucks), it has recently been noticed that the effectiveness of any vehicle wheel kinematics modifiers becomes reduced. There remains a significant need to address the wheel load path as it contacts the backup structure at unfavorable angles presenting a risk for increased intrusion in the dash area and the lower compartment negatively affecting the IIHS SORB structural and overall rating performance. In particular, there remains a continuing and significant need to provide additional improved impact performance in a vehicle that will include reducing intrusion of the forward structures, including in particular the wheel and tire of the vehicle, into the body or occupant compartment of the vehicle.


A method in a network node that includes a host and an accelerator, includes holding a work queue that stores work elements, a notifications queue that stores notifications of the work elements, and control indices for adding and removing the work elements and the notifications to and from the work queue and the notifications queue, respectively. The notifications queue resides on the accelerator, and at least some of the control indices reside on the host. Messages are exchanged between a network and the network node using the work queue, the notifications queue and the control indices.

In some communication networks, a network node processes data received over the network using a local accelerator. Various methods for delivering data to the accelerator are known in the art. For example, PCT International Publication WO 2013/180691, whose disclosure is incorporated herein by reference, describes devices coupled via one or more interconnects. In one embodiment, a Network Interface Card (NIC), such as a Remote Direct Memory Access (RDMA) capable NIC, transfers data directly into or out of the memory of a peer device that is coupled to the NIC via one or more interconnects, bypassing a host computing and processing unit, a main system memory or both.
PCT International Publication WO 2013/136355, whose disclosure is incorporated herein by reference, describes a network node that performs parallel calculations on a multi-core GPU. The node comprises a host and a host memory on which a calculation application can be installed, a GPU with a GPU memory, a bus and a Network Interface Card (NIC). The NIC comprises means for receiving data from the GPU memory and metadata from the host over the bus, and for routing the data and metadata towards the network. The NIC further comprises means for receiving data from the network and for providing the data to the GPU memory over the bus. The NIC thus realizes a direct data path between the GPU memory and the network, without passing the data through the host memory.

Low-latency processing in a network node


A computer implemented method, apparatus, and computer usable program code for processing a class file. The class file is loaded onto a target device. A determination is made whether verification is enabled. Static constraints are checked in response to determining the verification is enabled. A determination is made whether subroutines are present in a set of methods within the class file. The subroutines are in-lined in response to determining that subroutines are present in the set of methods.

1. Field of the Invention
The present invention relates generally to an improved data processing system and in particular to a computer implemented method and apparatus for in-lining Java™ subroutines. (Java and all Java-based trademarks are trademarks of Sun Microsystems, Inc. in the United States, other countries, or both.) Still more particularly, the present invention relates to a computer implemented method, apparatus, and computer usable program code for in-lining Java subroutines on a target device at load time.
2. Description of the Related Art
Java is an object-oriented programming language and environment focusing on defining data as objects and the methods that may be applied to those objects. The Java virtual machine (JVM) is a virtual computer component that resides only in memory. The JVM allows Java programs to be executed on different platforms as opposed to only the one platform for which the code was compiled. The Java virtual machine prescribes support for subroutines. Subroutines are used by Java compilers when a section of code must be executed from two or more paths in a particular method. In particular, Java compilers typically use subroutines to implement constructs, such as the finally clause of a 'try . . . finally . . . ' construct. Subroutines are implemented using any number of bytecodes, such as JSR, JSR_W, RET, and WIDE RET.
Unfortunately, subroutines contribute a significant amount of complexity to a Java virtual machine implementation. Areas of complexity include bytecode verification, Just-In-Time (JIT) compilation, and particularly liveness mapping for garbage collection. Subroutines make the flow analysis required for precise identification of object references very difficult, or in some cases, impossible.
In many cases, there are additional problems because subroutines from the subset of Java may not be supported on all devices, and newer versions of Java may only support subroutines for backwards compatibility or may not support subroutines at all. These problems cause difficulty because legacy applications will continue to use subroutines for the foreseeable future. One solution requires the Java virtual machine to have full support for subroutines which requires extensive memory and processing requirements. In another solution, applications must be run through a preprocessor using a separate application authoring-time system which removes JSR, JSR_W, RET, and WIDE RET bytecodes and in-lines the subroutines into each place where the subroutines are called. This process requires an additional build step and is not acceptable for many editions of Java used by various organizations.

International Business Machines Corporation


A method for establishing a support environment for a program may include identifying one or more support environment features depended upon by the program to run on one or more nodes, selecting one or more support software components indicated to provide the features, loading the selected support components on the one or more nodes, configuring the selected support components based on the program, and causing the program and the selected support components to be executed on one or more nodes.

It is common for computer programs to need to be loaded simultaneously with one or more support components in order to function properly. For example, a program in the PHP scripting language may need to be loaded simultaneously with a language runtime for the PHP scripting language, and possibly other support components. Where such a program makes use of a feature of the PHP scripting language that first appeared in a particular version of the PHP scripting language, the loaded language runtime for the PHP scripting language must be recent enough to support this feature.


A continuous type media mill for effecting in a particulate-containing substance, a reduction in size of the particulate. The mill includes a support and an elongate vessel mounted on the support. The vessel has a generally cylindrical interior. An inlet port to the interior of the vessel is provided as is an outlet port for the particulate-containing substance. Comminuting media is provided in the interior of the vessel, the media having a tendency to move toward the outlet port with the particulate-containing substance flowing through the vessel. A rotor extends centrally in the cylindrical interior of the vessel and along a majority of the length thereof. The rotor has a diameter substantially less than the inner cylindrical surface of the vessel to define a space therebetween. The rotor includes a plurality of blades spaced axially along the length thereof for flinging the media radially outwardly in response to a rotation of a driven shaft for the rotor. Structure is provided in the vessel for effecting a movement of the comminuting media toward the inlet port to the vessel in response to a rotation of the driven shaft to counter the aforesaid tendency of the media migrating with the particulate-containing substance toward the outlet port.

Many devices are known for effecting a reduction in the size of a particulate in a particulate-containing substance. One such device is disclosed in U.S. Pat. No. 4,684,072, assigned to the same assignee as is the present invention. The aforesaid patent and the prior art listed therein will reveal various structures that have been employed over the years to effect an agitation of the comminuting media to effect the desired result. Each of these various structures provides varying degrees of sheer forces and impact forces for effecting the desired reduction in size of the particulate.
It is frequently desirable to cause particle size reduction of a solid contained in a liquid. Liquids are frequently wanted and desired in the total product system. For example, liquids tend to replace undesirable gases held by the particle. Further, more uniform particle size distribution is obtainable with liquid additions and a desired reaction or bond may even be created. However, in continuous type mills, that is, mills wherein a product is continuously introduced into the comminuting vessel at one end and the comminuted mixture or product is removed at the opposite end, a strong desire has existed for many years in providing apparatus which would quickly process the product flowing therethrough so as to increase the productivity of the mill. However, the length of time that the product remains in the mill is directly related to the particle size of the particulate desired in the output mixture or product. Further, considerable energy is consumed during the period of time that the mill is in operation. Thus, the cost of energy per unit of product can be greatly diminished if the amount of product being processed through the mill can be substantially increased. It has been discovered that the construction of the rotor has a direct influence on the efficiency of the mill.
In addition to the subject matter disclosed in U.S. Pat. No. 4,684,072 mentioned above, Applicant also wishes to make of record U.S. Pat. Nos. 4,582,266 and 4,746,069, all of which are assigned to the same assignee as is the present invention. Reference to the aforesaid three patents is to be incorporated herein by reference.
In instances where a particulate is contained within a liquid, as the liquid and particulate move through the mill, there is a tendency on the part of the liquid and particulate to carry the comminuting media along therewith toward the outlet from the mill. As a result, and eventually, a majority of the media will have migrated toward and congregated around the outlet from the mill thereby making the mill operate inefficiently and consume a great deal of energy.
Accordingly, it is an object of this invention to provide a mill for continuously processing product, wherein the comminuting media is sufficiently agitated and with a minimum of friction occurring between the moving parts to achieve the desired amount of comminution of a particulate-containing product and without effecting a migration of the media toward the outlet end of the comminuting vessel.
It is a further object of the invention to provide a continuous type mill, namely, a mill wherein product is introduced at one end of the vessel and the comminuted mixture or product removed at the opposite end, wherein the comminuting media is sufficiently agitated and with a minimum of friction occurring between the moving parts to achieve the desired amount of comminution of a particulate-containing product and without effecting a migration of the media toward the outlet end of the comminuting vessel.
It is a further object of the invention to provide a rotor having structure thereon which causes the media to move upstream against the flow of the liquid and particulate moving through the comminuting vessel to counter the tendency of the liquid and particulate to carry the media toward the outlet end of the vessel.
It is a further object of the invention to provide a continuous type mill wherein the media is maintained evenly distributed throughout the length of the comminuting vessel even when liquid and particulate is flowing from the inlet end toward the outlet end.
It is a further object of this invention to provide a mill, as aforesaid, wherein an increase in the amount of the comminuted mixture or product is effected while simultaneously reducing the energy consumed in comminuting the product per unit of product produced.
It is a further object of this invention to provide a mill, as aforesaid, which is durably constructed and which will provide a generally maintenance-free operating characteristic.


In a semiconductor device in which transistors are formed in a plurality of layers to form a stack structure, a method for manufacturing the semiconductor device formed by controlling the threshold voltage of the transistors formed in the layers selectively is provided. Further, a method for manufacturing the semiconductor device by which oxygen supplying treatment is effectively performed is provided. First oxygen supplying treatment is performed on a first oxide semiconductor film including a first channel formation region of a transistor in the lower layer. Then, an interlayer insulating film including an opening which is formed so that the first channel formation region is exposed is formed over the first oxide semiconductor film and second oxygen supplying treatment is performed on a second oxide semiconductor film including a second channel formation region over the interlayer insulating film and the exposed first channel formation region.

1. Field of the Invention
The present invention relates to a method for manufacturing a semiconductor device.
In this specification, a semiconductor device generally means a device which can function by utilizing semiconductor characteristics, and an electrooptic device, a semiconductor circuit, and electronic equipment are all semiconductor devices.
2. Description of the Related Art
A technique by which a transistor (also referred to as a thin film transistor (TFT)) is formed using a semiconductor film formed over a substrate as a channel formation region has attracted attention. The transistor is applied to a wide range of electronic devices such as an integrated circuit (IC) or an image display device (display device). A silicon-based semiconductor material is widely known as a material for a semiconductor film applicable to a transistor. As another material, an oxide semiconductor has been attracting attention.
For example, a transistor including a semiconductor film formed using an amorphous oxide containing indium (In), gallium (Ga), and zinc (Zn) (In—Ga—Zn—O-based amorphous oxide) is disclosed (see Patent Document 1).
When such transistors each including an oxide semiconductor are stacked to have a three-dimensional layered structure, a complex circuit configuration can be formed in a small area. Further, by combination of an integrated circuit formed using a stack of such transistors each including an oxide semiconductor and a transistor which has high field effect mobility and formed using a single crystal silicon substrate, a higher functional circuit can be obtained.

Method for manufacturing semiconductor device

Semiconductor Energy Laboratory Co., Ltd.


A back-lighted clap board or slate used with the identification of rolls, takes and scenes and like information in the making of films that is back-lit with an electroluminescent panel. In addition, a liquid crystal display is formed on the clap board in front of the electroluminescent panel so that the liquid crystal display is back-lit and made visible and thus in dark or semidark conditions will be energized and back-lit with the light from the electroluminescent panel.

1. Field of the Invention
The present invention relates in general to electroluminescent panels and specifically relates to a self-contained electroluminescent back-lit clap board/slate or reusable writing board or surface that is used for the introduction of rolls, takes and scenes in the making of movies and the video films and which includes a liquid crystal clock display that is back-lit by the electroluminescent panel.
2. Description of Related Art Including Information Disclosed Under 37 CFR 1.97 and 1.98
In the making of movies and video films, where various rolls, takes, and scenes are involved, a camera assistant typically marks a slate board with a dry-erase pen to indicate the roll number, scene number, take number and other important information. The assistant, in such case, marks the clap board/slate with the appropriate information and must open the moveable arm of the clap board/slate, call attention to the operation that is about to take place, and force the moveable arm against a stationary arm to create an audio cue (soundtrack) for matching the visual cue (film/video) for editing purposes to synchronize soundtracks with scenes, takes, and the like as recorded on the clap board/slate surface. Very often the camera assistant is using such slate in a low-light or no-light situation. In the low-light or no-light situation, a slate light has been used in the past. This is a small light that requires batteries or alternating current and an operator who must try to hold the light on the clap board while it is being operated by the camera assistant. Other solutions to the problem have been the use of small flashlights by the operator. The operator attempts to keep the clap board illuminated with the flashlight or the small slate light while it is being held and operated by the camera assistant. This creates an obvious problem because the light does not evenly cover the slate and it is difficult to hold steady on the slate or clap board.
This problem was remedied in commonly assigned U.S. Pat. No. 5,457,507 which provided a self-contained electroluminescent back-lit clap board or slate or reusable writing board or surface made of transparent or semi-transparent plexiglass.TM., fiberglass or similar material that may be used with a dry-erase marking pen (overhead projector pen), fluorescent dry-erase marking pen or other nonpermanent writing utensils.
However, another important requirement that has been added to such clap boards is a time code area, the Society of Motion Pictures and Television Engineers (SMPTE) code for film and video production. The prior art clap boards utilize a high intensity one-inch tall LED readout. It has to be fed a continuous time code via cable or wireless signal.
Because one-inch tall LED devices are used for the readout, at least eight "C" batteries must be used. Clearly this creates a bulky package on the backside of the clap board and adds weight to the unit. Typical units weigh 3.3 pounds with the batteries.
It would be advantageous to have a clap board that has a time code area thereon that is lightweight and that requires low power and that would require very little space.

Self-contained electroluminescent back-lit clap board/slate having an LCD clock display


Methods of producing arylsulfatase A are described herein. The methods can include one or more steps of chromatography. Exemplary chromatographic steps include ion exchange chromatography, mixed mode chromatography, and hydrophobic interaction chromatography. Compositions containing and methods using arylsulfatase A produced by the methods described herein are also disclosed.

Metachromatic leukodystrophy (MLD) is caused by an autosomal recessive genetic defect in the lysosomal enzyme arylsulfatase A (ASA or ARSA), resulting in a progressive breakdown of membranes of the myelin sheath (demyelination) and accumulation of galactosyl sulphatide (cerebroside sulfate) in the white matter of both central nervous system (CNS) and peripheral nervous system. Patients with metachromatic leukodystrophy can be treated with enzyme replacement therapy (ERT) using recombinant arylsulfatase A (rASA).

Methods for purification of arylsulfatase A