Drug – bio-affecting and body treating compositions – Extract – body fluid – or cellular material of undetermined... – Blood
Reexamination Certificate
1998-08-24
2001-01-23
Saucier, Sandra E. (Department: 1651)
Drug, bio-affecting and body treating compositions
Extract, body fluid, or cellular material of undetermined...
Blood
C424S529000, C435S002000
Reexamination Certificate
active
06177106
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention relates to methods for fractionating red blood cells (RBC) of human blood into several fractions having different functions. Further the present invention relates to specific materials produced by such a method, which are possessed of antibacterial properties or inhibitory against bacterial proliferation.
Although it has been conventionally known that red blood cells of human blood act as carriers for carrying a large amount of oxygen and carbon dioxide at a high speed, the other functions of red blood cells have not yet been known completely. Notably, of the four signs of inflammation, rubor, calor, tumor, and dolor, the first three are caused by the congregation of red blood cells. The red blood cells that have gathered at the inflammatory site are estimated to have some action on the inflammation. However, in the past thirty years, there have been very few publications in this field.
A question pondered during the development of the present invention is whether the red blood cell truly is a single cell. The literature on the subject generally indicates that the red blood cell is a single cell. An experiment performed during development of the present invention, however, raised some doubt that the red blood cell is a single cell.
As part of the experiment, a drop of blood from a patient's cubital vein is extracted and then deposited into Costar's flask containing RPMI-1640. The contents of Costar's flask then is mixed.
FIG. 1
illustrates an example of the resulting mixture. The mixture then is exposed to twenty minutes of sunlight which, in turn, produces the results illustrated in FIG.
2
. The mixture then is incubated in a 5% CO
2
, 37° C. incubator.
FIG. 3
illustrates the condition of the mixture after 3.5 hours and
FIG. 4
shows the condition of the mixture after 24 hours. For purposes of comparison,
FIG. 5
shows an unprocessed mixture which is 24 hours old.
As
FIG. 2
demonstrates, the cells change in shape and nature when exposed to 20 minutes of sunlight. Nevertheless, after twelve hours of incubation, many of the cells will recover to their original condition. Some cells, however, are unable to recover fully. After 24 hours, the so-called “ghost cells” appear.
Next, synovial fluid is extracted from patients with arthrosis deformans of the knee, rheumatoid arthritis, and the like, and a drop of blood of the same type as the patient's is added thereto. Also added to the synovial fluid is RPMI-1640 in a quantity of about 3 milliliters.
FIG. 6
illustrates how the cells react. The way the red blood cells change is different depending on whether the red blood cells are those of patients suffering from arthrosis deformans of the knee or whether they are from patients afflicted with Rheumatoid arthritis.
The red blood cells therefore can be categorized into several types. Exposure to sunlight reveals five different types. The synovial fluid reveals six different types. Thus, there is reason to be skeptical about the notion that the red blood cell is a single cell.
SUMMARY OF THE INVENTION
It is therefore a primary object of this invention to provide a method of categorizing and extracting red blood cells.
It is another object of this invention to find new functions of red blood cells which are greater parts of human blood.
A still more specific object of this invention is to provide methods for fractionating red blood cells into different functions, which method can be used for, among other things, such research.
In order to achieve the above objects, the inventor of the present invention has performed research on the concept of fractionating red blood cells of human blood into several fractions. The inventor found that the tested blood could be fractionated into three layers after the blood was combined with dextran and then maintained under a certain condition for a certain period. The fractionated blood cells contained in these three layers provided different functions on bacteria, respectively. The present invention is based on this knowledge.
The above described objects are accomplished by the method for fractionating red blood cells of human blood into three fractions according to the present invention which comprises the following steps: (a) a human blood sample is mixed with dextran aqueous solution and is kept stationary for 60 to 75 minutes to fractionate this blood sample into three layers; (b) these upper, intermediate, and lower layers are individually separated and collected; and (c) the upper layer sample is treated with a hypotonic solution for a short period and then combined with a hypertonic solution.
Further, the present invention may provide another method for producing a fraction including antibacterial red blood cells, which comprises the following steps: (a) a human blood sample is mixed with dextran aqueous solution and is kept stationary for 60 to 75 minutes to fractionate this blood sample into three layers; (b) the upper layer is separated from the other layers; and (c) the upper layer is treated with a hypotonic solution for a short period and then is combined with a hypertonic solution.
Furthermore, the present invention may provide still another method for producing a fraction including bacterial proliferation inhibitory red blood cells, which comprises the following steps: (a) a human blood sample is mixed with dextran aqueous solution and is kept stationary for 60 to 75 minutes to fractionate this blood sample into three layers; and (b) the intermediate layer sample is separated and collected from the other layers.
Accordingly, the methods provided by the present invention can easily fractionate three different blood fractions including red blood cells as a main component and having different functions. These three different fractions may be applied to several clinical tests such as antibacterial tests and the like.
The present invention also provides a method for fractionating red blood cells into three layers, each of which includes a different type of red blood cells. The red blood cells in the upper layer are the upper layer (UL) RC, those of the intermediate layer are the intermediate layer (IL) RC, and those of the lower layer are the lower layer (LL) RC.
These and other objects and many of the attendant advantages of this invention will be readily appreciated as the same becomes better understood by reference to the following detailed description when considered in connection with the accompanying drawing.
Liniak, Berenato, Longarce & White, LLC
Saucier Sandra E.
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