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G01N35/ Devices for transferring samples or any liquids to, in, or from, the analysis apparatus, e.g. suction devices, injection devices characterised by the. Publication number, DE T2. Publication type, Grant. Application number, DE PCT number, PCT/US/ Publication date, 6 Mar. AAV-Vektoren, die Transgene tragen, wurden zB in Kaplitt et al. AAV vectors carrying transgenes, such as were in Kaplitt et al. () Nature Genetics.
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Diese Erfindung betrifft im Allgemeinen die Partikelanalyse. This invention relates to particle analysis in general. More specifically, it relates to methods and devices for performing automated blood cell analysis by "Impedance Analysis", "light scattering analysis" and "fluorescence analysis" Flow cytometry and are combined to form a whole.
Diese Erfindung betrifft auch ein Mehrzweckreagenzsystem und ein Verfahren zur schnellen Analyse einer Vollblutprobe. This invention also relates to a multipurpose reagent system and a method for rapid analysis of a whole blood
was ist der ungefähre Durchmesser der reifen Elternzelle?. Peripheral blood of a human usually contains red blood cells RBCplatelets PLT and white blood cells WBCall of which are suspended in a conductive medium commonly known as plasma.
Plasma umfasst Proteine, Anionen und Kationen. Plasma comprises proteins, anions and cations. Plasma also contains components that support the formation of blood clots. The blood of an adult usually contains about 4. Mature RBCs have no nuclei and are generally in the shape of circular biconcave disks with a diameter of about 7. RBCs contain hemoglobin which gives blood its red color. Hemoglobin helps transport oxygen and carbon dioxide and plays a role in maintaining pH in blood.
The blood of an adult usually contains abouttoplatelets per cubic meter. Platelets are small, biconvex cellular particles. Their general configuration includes a granular central portion embedded in a homogeneous matrix. Peripheral blood also contains red cells of earlier maturation levels which are important diagnostic indicators. Two of these are reticulocytes and nucleated red blood cells.
In the earliest stage of development the red cell consists was ist der ungefähre Durchmesser der reifen Elternzelle? of core and will be referred to as an erythroblast. When the erythroblast matures, the nucleus becomes smaller, seedless and increasingly nearly spherical. Consequently, the maturation includes a complete loss of the core. The immature red cells that retain a nucleus are called nucleated red blood cells NRBC.
The NRBC count has been useful in patient monitoring in many disease states. However, NRBCs in peripheral blood often contribute to inaccurate count of the white cell count, partly because of the presence of a nucleus which makes them difficult to distinguish from small white cells. Reticulocytes provide a means for evaluating the anemic condition of a patient. Anemia usually occurs as a result of an unbalanced increase in the rate of removal of erythrocytes from blood, or a reduction in the rate at which they are formed and released into the blood.
An increased Retikulozytenpatientenzahl in an anemic patient indicates rapid erythrocytes turnover which suggests acute blood loss or hemolysis. In normal human blood, the concentration of white blood cells, called leukocytes or WBCs, much lower than the concentration of red cells.
The normal concentration of WBCs is approximately per microliter. They vary in size, most of about 7. They are more nearly spherical in shape than RBCs and usually somewhat larger the volume.
WBCs can be generally classified as either granular or non-granular. The granular WBCs include Neutrophite, eosinophils and basophils. The non-granular WBCs include monocytes and lymphocytes. These categories of WBCs are often referred to collectively as a "five-part differential," and in general, the most significant of these categories neutrophils and lymphocytes.
Their cytoplasm contains numerous tiny granules which can be stained. Under certain conditions neutrophils may leave the blood vessels and dissolve, which granules are released into the connective tissues. These granules are rich in certain enzymes that are active and participate in the defense mechanism of the body.
The core of a normal lymphocyte occupies nearly the entire cell volume, and thus the cytoplasm surrounding the nucleus, a rather thin shell. Lymphozytenzytoplasma can be stained with dyes due to content of ribonucleic acid from Zytomplasma. Lymphocytes may leave the blood vessels and enter the connective tissue where they also constitute a part of the defense mechanism of the body, where they play a major role in the immune response of the body.
Es gibt drei Haupt-"teilmengen" von Lymphozyten, die was ist der ungefähre Durchmesser der reifen Elternzelle? klinisch bedeutend sind: There are three main "subsets" of lymphocytes that are currently clinically significant: T lymphocytes, B lymphocytes and natural killer Zeller, also known as "large granular lymphocytes" or NK cells.
Each of these subsets can be based on the existence of distinguished characteristic cell surface markers or antigens. Also have during T-lymphocytes have a high density of immunoglobulin of their surfaces, little or none of B-lymphocytes. T-lymphocytes are characterized by various surface markers against which antibodies can be formed. Categories of T lymphocytes have been identified according to their surface markers and overall function. The "helper T cells" help B cells produce certain classes of antibody molecules, and help other T cells in their immune response.
The "suppressor T cells" are regulatory cells that can suppress the responsiveness of other T or B cells. The suppressor T cells include several subsets which are also recognized by different surface markers.
The ability to count blood cells to classify classify, is useful for assessing the health of an individual. For example, the levels of circulating CD4 lymphocytes helper-T cells having a CD4 antigen expressed on the surface of the cell is currently regarded as the best single predictor for the progression of HIV infection. The CD4 levels can be used to classify individuals for inclusion in experimental treatment systems, to determine when to start with an antiviral therapy, and to monitor the treatment response in clinical trials.
Because CD4 lymphocyte levels for some HIV-infected individuals may be important, it is desirable to measure this parameter accurately. According to the current state of the art of cell analysis, two techniques for counting and classifying cells are used. Diese sind allgemein als "Durchflusszytometrie" und "Bildzytometrie" bekannt. These are commonly known as "flow cytometry" and known "image cytometry.
The Durchflusszytometrietechnik which is substantially that cells each having a are passed through a measuring zone of a flow cell according to the other, is preferred in clinical applications where patient test load is an important measure. Mainly is because it has at least one order of magnitude advantage in the number of cells that can be analyzed per second. The instrumentation incorporates flow cytometry can be further subdivided into two methods which can be generally classified as "conventional hematology" and "Fluoreszenzzytometrie".
Deshalb kann das Fluoreszenzverfahren verwendet werden, um was ist der ungefähre Durchmesser der reifen Elternzelle? Zelltypen in feinere Klassifikationen zu unterteilen.
Therefore, the fluorescence method can be used to subdivide the cell types into finer classifications. A second distinction between the two methods is that conventional hematology gives results in absolute terms, while Fluoreszenzzytometrieergebnisse present in relative terms. Hematology analyzers provide accurate volume and dilutions and are therefore able to measure absolute cell concentrations, or absolute numbers of cell types per microliter of human blood.
Das Fluoreszenzzytometrieverfahren ergibt nur relative Konzentrationen oder prozentuale Anteile der verschiedenen Zelltypen. The Fluoreszenzzytometrieverfahren gives only relative concentrations, or percentages of the different cell types.
A third distinction is that the hematology method is generally automated, the Fluoreszenzzytometrieverfahren, as it is commonly listed practically today is, at best,
was ist der ungefähre Durchmesser der reifen Elternzelle?, both in sample preparation as well as in the sample analysis. The Fluoreszenzzytometrieverfahren is therefore significantly more labor intensive than the hematology method.
Beide Verfahren benutzen die Zelle bei Zelle-Analyse. Both methods use the cell at cell analysis. Therefore, it is necessary because of the high concentration of cells in whole blood to dilute the blood samples prior to analysis so that individual cells can be isolated for sensing within a flowcell. Patientenzahlen sind allgemein definiert als die Zahl der "Ereignisse" pro Mikroliter Blut.
Was ist der ungefähre Durchmesser der reifen Elternzelle? numbers are generally defined as the number of "events" per microliter of blood.
Each of these devices uses combinations of scatter, impedance, and absorption to distinguish and quantify cells, and can therefore be classified as a conventional hematology instrument. Im Gegensatz dazu sind im Fluoreszenzdurchflusszytometer die Prinzipien von Fluoreszenzzellanalyse mit Lichtstreuung zusammengefasst. In contrast, the principles of fluorescence cell analysis are summarized with light scattering in Fluoreszenzdurchflusszytometer.
In general, this requires that the cell is stained with a suitable dye, or that a fluorochrome label be attached to an antigen or antibody on the cell's surface thus indicating the occurrence of a specific antigen-antibody reaction.
In the Fluoreszenzdurchflusszytometrie a suspension of previously stained or fluorescently labeled particles, typically cells in a blood or other biological liquid sample transported through a flowcell where the individual particles in the sample with one or more focused light beams are illuminated. One or more detectors detect the interaction between the light rays and the labeled particles flowing through the flow cell.
Usually some of was ist der ungefähre Durchmesser der reifen Elternzelle? detectors are designed to measure fluorescent emissions, while other detectors measure scatter intensity or pulse duration.
Consequently, can be mapped into a feature space, each particle passing through the flow cell, whose axes are the emission colors, light intensities, or other properties, namely scattering are measured by the detectors.
Preferably, the different particles can be mapped into distinct and non-overlapping regions of the feature space in the sample, which makes it possible that each particle is analyzed based on its mapping in the feature space. In this respect, flow cytometry differs from the conventional hematology instruments in that some of the feature space axes fluorescence emissions include. Wie oben angemerkt sind Lymphozytensubklassen entscheidende Gesundheitsfaktoren. As noted above, lymphocyte subclasses are key determinants of health.
Therefore, it is desirable that these and other parameters are measured precisely. An example of this problem is the CD4 cell patient count. The following articles discuss additional difficulties with developing CD4 patient counts using current methods and devices: One reason for
was ist der ungefähre Durchmesser der reifen Elternzelle? in CD4 patient numbers is the sample preparation by hand, which can not be accurately regulated and is dependent on the skill of the operator.
For example, a conventional process starts for determining a CD4 patient with the removal of two tubes of blood from a patient.
The sample preparation steps for the Durchflusszytometrietests are labor intensive and operator dependent. These steps do not readily lend themselves to automation and precision. A volume of the desired fluorochrome labeled monoclonal antibody is added. Choosing the right time during the Lysestadiums is important. If the operator does not allow the lysis reaction can continue long enough, RBCs may remain in the sample and distort the measurements.
If the operator permits the lysis reaction is continued too long, the lyse may attack the WBCs. After determining that the lysis reaction is complete, the operator the sample was centrifuged and washed to remove any debris left over was ist der ungefähre Durchmesser der reifen Elternzelle? lysed RBCs.
Fragments, red cell "stroma" can interfere with the detection method of the typical flow cytometer.
Use of first and second compositions for the manufacture of a medicament for the treatment of Parkinson's disease, wherein the first composition comprises recombinant adeno-associated virus rAAV virions comprising one transgene encoding an aromatic amino acid decarboxylase AADC , and a pharmaceutically acceptable excipient to be administered to the cranial cavity, and wherein the second composition comprises L-dopa and a pharmaceutically acceptable excipient for oral use, wherein only one transgene is administered.
Use according to claim 1, wherein the administration is a convection-enhanced delivery CED. Using first and second therapeutic compounds in the manufacture of first and second medicaments for use in a method for the treatment of Parkinson's Disease, wherein the first compound rAAV virions comprising recombinant adeno-associated virus comprising a transgene encoding an aromatic amino acid decarboxylase AADC for delivery to the cranial cavity, and wherein the second compound comprises L-dopa for oral use, wherein only one transgene is to be administered.
Use according to claim 3, wherein the administration to the cranial cavity via convection-enhanced delivery using an osmotic pump or infusion occurs. Use according to any of claims 2 to 4, wherein the administration to the striatum occurs. Gebiet der Erfindung Field of the Invention. The present invention relates generally to efficient delivery of viral vectors to the CNS. In particular, the present invention relates to gene therapy for the treatment of disorders of the central nervous system CNS , particularly those disorders which involve the neurotransmitter dopamine.
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