Translate

Thursday, 25 December 2014

Thrombopoietin (TPO)

A human thrombopoietin (TPO) is a 332 amino acid, 60 kDa glycoprotein, containing six potential N-linked glycosylation sites. These are all localized towards the C terminus of the molecule. The N-terminal half exhibits a high degree of amino acid homology with EPO and represents the biologically active domain of the molecule.

TPO is the haemopoietic growth factor now shown to be the primary physiological regulator of platelet production. This molecule may, therefore, represent an important future therapeutic agent in combating thrombocytopenia, a condition characterized by reduced blood platelet levels. The most likely initial TPO therapeutic target is thrombocytopenia induced by cancer chemo- or radio therapy. This indication generally accounts for up to 80 per cent of all platelet transfusions undertaken. In the USA alone, close to 2 million people receive platelet transfusions annually.





Platelets (thrombocytes) carry out several functions in the body, all of which relate to the arrest of bleeding. They are disc-shaped structures 12 μm in diameter, and are present in the blood of healthy individuals. They are formed by a lineage-specific stem cell differentiation process. The terminal stages of this process entail the maturation of large progenitor cells termed ‘megakaryocytes’. Platelets represent small vesicles that bud off from the megakaryocyte cell surface and enter the circulation.

Monday, 20 October 2014

Leukocytes, their Range & Function

Leukocytes (white blood cells) encompass all blood cells that contain a nucleus, and these cells basically constitute the cells of the immune system. They thus function to protect the body by inactivating and destroying foreign agents. Certain leukocytes are also capable of recognizing and destroying altered body cells, such as cancer cells. Most are not confined exclusively to blood, but can circulate/exchange between blood, lymph and body tissues. This renders them more functionally effective by facilitating migration and congregation at a site of infection.

Leukocytes have been subclassified into three families: mononuclear phagocytes, lymphocytes and granulocytes. These can be differentiated from each other on the basis of their interaction with a dye known as Romanowsky stain.

Mononuclear phagocytes consist of monocytes and macrophages, and execute their defence function primarily by phagocytosis. Like all leukocytes, they are ultimately derived from bone marrow stem cells. Some such stem cells differentiate into monocytes, which enter the bloodstream from the bone marrow. From there, they migrate into most tissues in the body, where they settle and differentiate (mature) to become macrophages (sometimes called histocytes). Macrophages are found in all organs and connective tissue. They are given different names, depending upon in which organ they are located (hepatic macrophages are called Kupffer cells, central nervous system macrophages are called microglia, and lung macrophages are termed alveolar macrophages). All macrophages are effective scavenger cells, engulfing and destroying (by phagocytosis) any foreign substances they encounter. They also play an important role in other aspects of immunity by producing cytokines, and acting as antigen-presenting cells.


MONOCYTES


Lymphocytes are responsible for the specificity of the immune response. They are the only immune cells that recognize and respond to specific antigens, due to the presence on their surface of high-affinity receptors. In addition to blood, lymphocytes are present in high numbers in the spleen and thymus. They may be sub-categorized into antibody-producing B-lymphocytes, T-lymphocytes (which are involved in cell-mediated immunity) and null cells.



T-lymphocytes may be subcategorized on a functional basis into T-helper, T-cytoxic and T-suppressor cells. T-helper cells can produce various cytokines which can stimulate and regulate the immune response. T-cytotoxic cells can induce the lysis of cells exhibiting foreign antigen on their surface. As such, their major target cells are body cells infected by viruses or other intracellular pathogens (e.g. some protozoa). T-suppressor cells function to dampen or suppress an activated immune response, thus functioning as an important ‘off’ switch.



Most T-helper cells express a membrane protein termed CD4 on their surface. Most T-cytotoxic and T-suppressor cells produce a different cell surface protein, termed CD8. Monoclonal antibodies specifically recognizing CD4 or CD8 proteins can thus be used to differentiate between some T cell types.





Null cells are also known as ‘large granular lymphocytes’, but are best known as ‘natural killer’ (NK) cells. These represent a third lymphocyte subgroup. They are capable of directly lysing cancer cells and virally infected cells.



GRANULOCYTES


The third leukocyte cell type is termed granulocytes, due to the presence of large granules in their cytoplasm. Granulocytes, many of which can be activated by cytokines, play a direct role in immunity, and also in inflammation. Granulocytes can be subdivided into three cell types of which neutrophils (also known as polymorphonuclear leukocytes; PMN leukocytes) are the most abundant. Attracted to the site of infection, they mediate acute inflammation and phagocytose opsonized antigen efficiently due to the presence of an IgG Fc receptor on their surface. Eosinophils display a cell surface IgE receptor and, thus, seem to specialize in destroying foreign substances that specifi cally elicit an IgE response (e.g. parasitic worms). These cells also play a direct role in allergic reactions. Basophils also express IgE receptors. Binding of antigen–IgE complex prompts these cells to secrete their granule contents, which mediate hypersensitivity reactions.

Wednesday, 21 May 2014

Biochip

A biochip is a device that has some of the features of a computer chip but, instead of doing calculations, it uses living cells or molecules from living cells to greatly speed up certain laboratory tests. A typical biochip is a glass or plastic chip or tile a few inches on a side. It has hundreds or even tens of thousands of microscopic droplets of material stuck to its surface like gum on a sidewalk. A computer looks at the chip using a camera. Information from a biochip can be used to learn about the differences between genes, cells, or drugs. It can also be used to study many other questions about cells. Biochips are also called microarrays, where micro means ‘‘small’’ and an array is any regular grid, such as a chessboard. The droplets on a biochip are laid down in a checkerboard pattern. A square chip five inches on a side may have 40,000 or more spots on its surface.

The most common kind of biochip is the DNA microarray, also called a gene chip or DNA chip. Deoxyribonucleic acid (DNA) is the long, coded molecule used by all living things to pass on traits to offspring. DNA also tells each cell how to make all the molecules it needs to live, like a cookbook containing many recipes. The DNA of almost every living thing is at least slightly different from that of every other.

In one type of DNA chip, genes (short pieces of DNA that code for single molecules) are placed on the chip. Since even large molecules are too small to see with the naked eye, millions of copies of each gene can be placed on a tiny spot on the chip.



HOW DNA CHIP WORKS?




In a DNA chip, each separate spot (also called a probe) contains one type of defective gene. To find out if a person has any of these defective genes in their own DNA, DNA is taken from the person’s cells. Copies of the person’s DNA are made, and these copies are labeled, meaning that they include a chemical that glows when ultraviolet light (which is invisible to the eye) shines on it. Small drops of liquid containing labeled copies of the person’s DNA are then added to the spots on the biochip.

A normal DNA molecule is shaped like a ladder, but the DNA copies being mixed on the biochip are one-sided copies, like a ladder that has been sawed in half lengthwise, cutting every rung in half. When two pieces of one-sided DNA that have matching rungs (or bases, as they are called) meet, they lock or zip together. When this happens, the two pieces of DNA are said to hybridize. If the patient’s genes match any of the defective genes that have been put on the biochip, they will attach to (hybridize with) those defective genes.

The chip is then washed to remove any of the person’s DNA that has not found a match on the chip. Finally, the chip is placed in ultraviolet light, and a camera records any spots that glow. These are spots where the labeled copies of the person’s DNA have matched up with DNA on the chip.

Examining a patient’s DNA for defects is called genetic screening. By using a biochip, genetic screening can be done very quickly, all the tests can be done at once, rather than doing hundreds or even thousands of separate tests.

Genetic screening is only one way of using biochips. Another important use for biochips is to study how genes are used by living cells. Each gene tells the cell how to make a certain protein molecule. Cells read the recipe given by the gene by first making another molecule, mRNA, which copies the information in the gene. The mRNA can then go to a place in a cell that will build the molecule that the gene codes for. The more mRNA a cell has for a gene at a particular time, the more it is said to be ‘‘expressing’’ that gene, i.e, the more of that particular molecule it is making. Gene expression changes all the time for thousands of genes in every cell.

In the laboratory, scientists can make DNA molecules from the mRNA found in a cell. This matching DNA is called cDNA (complementary DNA). If a biochip has all the genes of an organism dotted on its surface, then cDNA made from the mRNA in a cell can attach to (hybridize with) the genes on the chip. The more a gene is being expressed in the cell, the more cDNA for that gene there will be, and the more that cDNA will stick to the matching genes on the biochip. Spots with more labeled cDNA will glow more brightly under ultraviolet light. In this way, scientists can literally take a snapshot of how the genes in a cell are being expressed at any one time—how much the cell is making, at that moment, of thousands of different substances. This is extremely useful in trying to understand how cancer cells grow and in many other medical problems.