Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving fixed or stabilized – nonliving microorganism,...
Reexamination Certificate
1998-03-03
2002-10-01
Chin, Christopher L. (Department: 1641)
Chemistry: molecular biology and microbiology
Measuring or testing process involving enzymes or...
Involving fixed or stabilized, nonliving microorganism,...
C435S040500, C436S063000, C600S562000
Reexamination Certificate
active
06458555
ABSTRACT:
BACKGROUND OF THE INVENTION
The present invention resides in a method of examining the mucous membranes of the gastro-intestinal system, the urinary tract and the bronchial system.
All the knowledge concerning the mucous membranes available to date has been gathered by the so-called histological method—a method of studying tissues developed by Virchow—wherein tissue is technically prepared and treated with formaldehyde. This method provides for an excellent view of the various tissue structures and facilitates a safe diagnosis on the basis of changes in the tissue structure. An important disadvantage of this method, however, is that a particular layer at the surface of the mucous membrane is lost during the technical preparation of the tissue by the histological method: By exposure to formaldehyde, the mucous coagulates and is washed away by the various solutions used during the further method steps. As a result, histological studies do not involve the particular layer at the surface of the membrane and no appropriately accurate knowledge is obtained concerning the microbiology present in this layer. Important aspects concerning this layer have therefore never come to the attention of scientists.
Another method for examining the mucous membranes resides it the so-called brush cytology. With this method, a small metal brush is introduced through an endoscope in contact with the mucous membrane surface and the membrane is streaked by the brush. However, this kind of tissue removal leads to bleeding of the injured mucous membrane so that a substantial amount of blood is mixed with a relatively small amount of tissue obtained with this method. The tissue therefore, must first be cleaned by centrifuging, whereafter only a few cells remain for studying and examination. A thorough examination or a study of the mucous-epithelial layer or of the microbiology of this layer is impossible with this method.
It is the object of the present invention to provide a method by which mucous membranes can be studied and examined while the microorganisms living on, or in, the mucous membrane are not only not destroyed, but are clearly recognizable.
SUMMARY OF THE INVENTION
In a method for the cytological examination of mucous membranes, a mucous particle of about pinhead size is removed from the surface of the mucous membrane and is moved across a slide surface. In this way, viscous mucous and, together therewith, the epithelial surface layer portion remain attached to the slide surface so as to form a streak preparation which is then dried for examination under a microscope. For moving the particle across the surface a needle is used which is placed with its tip onto the surface at a distance from the particle so that any pressure applied to the needle is directly transmitted to the slide surface. The needle is held at an angle of 10-30° so that the particle is contacted by the needle above its center such that a downward force is generated on the particle by which the particle is engaged with the surface in a reproducible fashion.
The mucous membrane is a tough viscous mucous layer which firmly adheres to the epithelial layer—in contrast to the liquid mucous within the stomach. The mucous membrane serves as protection from the aggressive stomach acids. Below this layer, microorganisms find optimal living conditions. In fact, this layer protects the microorganisms from acids and various mechanical attacks.
With the method according to the invention, a certain layer for example of the mucous membrane of the stomach can be representatively displayed and it can be shown that it contains substantial microbiological life. It can be shown with this method, that not only
Helicobacter pylori
live on the surface of the mucous membrane, but that, in the mucous epithelial layer, there are large amounts of
H. pylori,
a large growth of fungi such as yeasts of the genus Candida, and also molds. With the method according to the invention, it was finally possible to determine the cause for the chronic gastritis and of the chronic colitis.
The method according to the invention is also suitable for studying the life cycle of those microorganisms or to answer other scientific questions concerning mucous membrane problems. Although, below, the mucous membranes of the stomach are considered, the mucous membranes of the colon, of the bladder and of the bronchial tubes can also be studied with the method according to the invention even though the circumstances are different. Since in these cases, the highly viscous membrane of the stomach is not present, the microorganisms live on, and in, the epithelial cell layer of the mucous membrane.
REFERENCES:
patent: 4759376 (1988-07-01), Stormby
patent: 4777133 (1988-10-01), Picciolo et al.
patent: 5178859 (1993-01-01), Simon et al.
patent: 5357977 (1994-10-01), Michels
Maile, Laboratory Medicine: Hematology. Saint Louis, C.V. Mosby Company. 1972, pp. 1207-1208.*
Debongnie et al., 1994. Touch Cytology: a quick, simple, sensitive screening test in the diagnosis of infections of the gastrointestinal mucosa. Archives of Pathology and Laboratory Medicine 118(11): 1115-1118.*
Debongnie et al., 1992. Cytology: a simple, rapid, sensitive method in the diagnosis ofHelicobacter pylori. Am. J. Gastroentero 87(1): 20-23.*
Faverly et al., 1990. Identification ofCampylobacter pyloriin gastric biopsy smears. Acta Cytologica 34(2): 205-210.*
DeFrancesco et al., 1993.Helicobacter pyloriin gastroduodenal diseases: rapid identification by endoscopic brush cytology Diagnostic Cytopathology 9(4): 430-433.
Bach Klaus J.
Chin Christopher L.
Grun James L.
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