Analysis methods of the blood

Immunological investigation

The first thing we did in the immunological investigation was to check for tumors by means of various tumor markers, such as those looked at in a prostate checkup. In addition, markers for Hepatitis A, B, and C, were examined. Once a week we could also do a hepatitis investigation as well as a titration test (concentration test) on many different antibodies.

In addition, during the immunological investigation, values were determined for free T3 and T4 hormones.

T3 and T4 are formed in the thyroid as iodine thyroid hormones, stored there in protein, and then delivered after stimulation by a hormone (TSH) into the bloodstream. In the blood, T3 and T4 are mostly bound to proteins. These hormones promote the metabolism, growth, physical and mental development and have many other effects on different bodily functions (see Lit.1).

The equipment for the investigation in the hospital laboratory was a "Hitachi Cobra Integra 400 plus". It is very sensitive and requires a very exact use. The cooling chain of the inspection samples should not be interrupted for very long and the reagents have to be regularly renewed. We helped with the daily calibration of the equipment and the refilling of the reagents.

Blood sedimentation

For blood sedimentation the blood is drawn into a long vacuum tube (vacuette) with black markings. The "Sed rate" equipment can then measure, in a well vibrated sample, the sedimentation of the blood particles very quickly. Another method for determining the blood sedimentation is the manual evaluation: The vacuette is placed onto a scale. After one hour, you read how far the firm components of the blood have dropped and then again, after another hour has passed.

Blood serum

One of the most important investigations was the investigation of blood serum. For this special investigation (centrifuging the sediment) it was very important to use the appropriate vacuettes. The serum tubes were provided with a gel, ficoll, which makes the separation from serum and sediment possible: Higher up in the tube, one finds lymphocytes and monocytes while below, the erythrocytes thrombocytes and granulocytes are found.

In our case the tubes were frozen in order to preserve the blood serum. After they were thawed, the samples were spun in a centrifuge at 5000U/m for 10 minutes, so that the blood sediments and serum would again separate. If the serum is pink or red it is hemolytic ie the erythrocytes were either damaged in the process of removing the blood, or there is a genetic condition known as hemolysis present.

Blood clotting

The blood's ability to clot is very important for the process of wound closure and healing. If humans possess too few platelets, like thrombocytes, then the wound cannot close fast enough or, in the most unfavorable case, not at all, resulting in the patient bleeding to death. The INR is an international value for the blood clotting.

Investigation by means of electrophoresis

electrophoresis the blood is examined by dying the protein components with albumin, α1, α2, β and γ. The equipment is fully automated - e.g. washing and a dabbing and the recognition of the size of the platelet. The platelet is then given a buffer for swelling. The only procedure, which is not automated, is the pipetting of the sample into its chamber. The entire process takes only about one hour until the results are ready.

Blood cell count

A piece of equipment named the "Sysmex SF-3000" examines the blood cell count. In the well vibrated blood the red and white blood cells as well as the blood platelets are counted. If after this test, deviating values which cannot be clearly explained, are diagnosed, a more exact investigation must be made by means of microscopic differentiating.

Differentiation by means of microscope

Differentiating is the micro copying of the blood cells. The leukocytes are divided by percentage into their individual cell types (granulocytes, lymphocytes and monocytes). In addition, a drop blood on a slide must be prepared, dried for 15 minutes, and dyed with a coloring agent. The first coloring step is to immerse the slide into a May-Green-Forest solution for 5 minutes. After one minute of flushing with distilled water, the slide is immersed a second time for 15 minutes into a Giemsa solution and afterwards rinsed with tap water. The colored slide is evaluated with the help of the microscope. Under the microscope one can differentiate between the different types of the leukocytes and/or count them, thus creating a quantitative evaluation.

Blood parameter

Liver values

GOT (AST)
UGOT stands for Glutamat Oxalacetat Transaminase, while AST stands for Aspartat Amino Transferase. GOT is an enzyme, which very frequently is found in the heart muscle, skeletal muscle and liver cells. If the liver becomes diseased, an increased GOT value is usually found. Very frequently, liver illnesses are liver inflammation, cirrhosis of the liver, liver poisoning, liver cancer as well as different illnesses of the gall bladder or bile. A too high GOT value can likewise indicate a heart infarct or various illnesses of the skeletal musculature. The GOT limit values normally lie between 10-38U/l*.

GPT (ALT)
GPT refers to Glutamat Pyrovat Transaminase, and is also known as ALAT or Alanine Amino Transferase. GPT occurs in very high concentrations in the liver cells, and in smaller concentrations in the skeletal and heart musculature. This value is very sensitive. An increased value can be found after even a single cell is damaged. A GPT value is considered normal if it is 10-41 units per liter*.

Gamma GT
Gamma GT is called also gamma Glutamyl Transferase. An enzyme, which when found at a higher than normal value leads to damage to liver cells and/or damage of the bile ducts. Gamma GT values are very sensitive and even a small deviation can signify serious damage. Too high gamma GT values are usually associated with hepatitis, fatty liver, liver cancer and pancreatic illnesses, and lastly with alcohol abuse. The reference value of gamma GT should lie between 10-71U/l*.

AP
AP stands for Alkaline Phosphatase. AP is very important for the bones, intestines, and kidneys. AP is an enzyme which occurs in all somatic cells. One finds deviations from the normal AP levels especially in infancy, during growth and pregnancy, however, such deviations carry no negative consequences. In addition, diseases such as hepatitis, jaundice, metastatic bone disease, alcohol and pharmaceutically caused liver damage can seriously increase AP-values. The standard values of AP are between 40 and 129U/l.*.

Blood fats

Cholesterol
Cholesterol is manufactured by the body or absorbed from food. It is an important component of cell walls and serves in the production of hormones and stomach acid. Cholesterol is a type of fat and does separate in water, or in blood. Lipoproteins enclose the cholesterol molecule, in order to make it transportable. All transportation of Cholesterol is done by means of Lipoproteins. Lipoproteins are divided into LDL (Low Density Lipoprotein) and HDL (High Density Lipoprotein).

LDL carries the cholesterol from the blood to the individual organs. Surplus cholesterol can become deposits within arteries and lead to arteriosclerosis. Arteriosclerosis causes the blood vessels to become clogged and can lead to a cardiac infarct or an apoplexy. The main causes of this clotting are genetic predominance or a lack of exercise. The cholesterol level for adults should not be higher than 200mg/dl*.

HDL functions as the opposite of LDL. HDL transports surplus cholesterol to the respective organs which creates a protection from the LDL transported cholesterol. HDL collects the surplus Cholesterol and brings it to the liver, where the cholesterol can be broken down and purged. HDL should always be present in some quantity, particularly since the body cannot have too much of it. The value is particularly high with athletes, because of their high level of exercise.

It is important however that the ratio of LDL to HDL maintain a quotient of at least 5 or below.

To improve the values of LDL and HDL a balanced and varied nutrition is extremely helpful. One does not have to do without fat, but products such as fish, oat bran, and fiber are good components of a healthy diet. Exercise also helps to increase the amount of HDL present in the body.

Triglycerides are long chains of fatty acids, which serve as a supplier of energy. Eating usually serves to increase the amount of triglyceride present in the body. Triglycerides often deposit themselves on the hips as "reserves" and also serve as thermal insulation. The value of the triglyceride should not exceed 200mg/dl*.

Sugar values and insulin

Insulin
The hormone insulin is refined in the pancreas as pro-insulin which is built out of insulin and C-Peptid. Insulin is very important to the functioning of the body. In combination with other hormones it regulates the blood sugar level. A lack of insulin leads too chronically increased blood sugar. The common disease Type One diabetes is caused by an insulin deficiency. Type Two diabetes is different. Sufficient insulin is produced, but cannot be used by the recipient cells. This is caused by continuous exposure to high insulin levels. A normal value for insulin should not exceed 20μU/ml*.

Kidney values
The kidneys are the decontamination organ of the body and clean the blood. They collect waste and convert it to urine. When muscle cells break down, creatine, a by-product of muscle degradation, is processed by the kidneys. A low creatine level is desirable. If the creatine level exceeds 1,1mg/dl*, a more thorough blood and urine investigation should be performed to determine if the kidneys are malfunctioning.

Electrolytes
Calcium is very important for bone metabolism and should always be present in the body. Generally, the body does not transport water venously rather as a different isotonic solution. This is because if exposed to normal water, the process of osmosis would cause the cell to absorb the water until bursting. Because of this the proper concentration of magnesium and calcium in and around the cell must be maintained.
The limit values for sodium should lie between 130 and 147mmol/l*, for calcium between 2,02 and 2,60mmol/l*, and for potassium 3,8 and 4,6mmol/l*.

Trace elements
Trace elements are essential to the body and since the body cannot produce them, they must be procured from the diet, since they are acquired in trace quantities from our food.They are nevertheless urgently needed by the body (usually for enzyme synthesis). With the trace elements both, lack of or a surplus of them, can lead to large problems.

A lack of trace elements causes a deficiency syndrom in organs, but this can usually be treated by restoring the element levels.

Iron values

Iron is important for the energy production of cells and the transportation of oxygen. Women and girls have generally less iron, due to the loss of blood through menstruation (every hemoglobin of human blood contains four iron ions). A lack of iron is determined by reviewing the hemoglobin and MCV levels. If both levels are lower than normal the patient has an iron deficiency, signs of which are pallor, tiredness and or cracked lips. The standard level of iron should lie between 59 and 158μg/dl*

An iron surplus is rather rare, and usually results from the illness hemochromatosis. This illness can present at age 30-40 and cause heart and liver damage. With iron surplus the ferritin levels must also be closely watched.

The protein ferritin indicates how much iron is stored in the body. A low ferritin level can mean a lack of iron and thus a lack of hemoglobin. Also, the ferritin level is different for women than for men, again as a result of menstruation. Ferritin levels are considered too low at 15ng/ml, but only if the other corresponding values (hemoglobin and MCV) are also low. A too low ferritin value with men could be the result of iron absorption caused by an intestinal problem. Here the limiting levels lie at approx. 25-30ng/ml*.

Transferrin is produced in the liver and is the main transportation protein for iron from food into the blood. Transferrin should be examined in direct connection with Ferritin and iron levels. That means that a deviation of transferrin should only be considered if there is a corresponding deviation in the iron level.

The transferrin saturation is a measure of how many molecules are actually currently occupied with the transport of iron. Too small a value, (<15%* , should be avoided as iron will not be able to be absorbed from meals even if present. If the iron level is too low, the body tries to take up more iron by increasing the production of transferrins.

Copper and caeruloplasmin

Copper and caeruloplasmin–130μg/dl*) has a great many uses within the body. There are no well known dangers from too high or low copper levels, and copper is not usually tested for with common bloodwork.

If there is a copper surplus, the body can separate the unnecessary copper. With a deficiency the cause should be examined particularly the level of caeruloplasm (16– 35 mg/dl*) present. This value is almost always maintained and will adjust itself in case of deviations. Since the role of the copper has not been thoroughly investigated we did test it in our research.

Inflammation values

Capsel Reactive Protein, CRP, is a protein, which arises with inflammations. Even with a low cold or a light injury, the CRP levels rise to greater than 0,5mg/dl*. This value should lower itself independently. However, it does not indicate a viral infection. If in a healthy patient the levels are high without a clear cause then this can be a result of an increased risk for Arteriosclerosis and consequently, an increased risk for cardiac infarct and/or apoplexy.

Blood cells and blood cell count

AAll blood cells are formed (hematopoiesis) in the bone marrow and from there distributed throughout the body. Under the microscope one can see many different blood cells.

The erythrocytes, or erys for short, are the red blood corpuscles and do not possess a cell nucleus. They are responsible for the energy metabolism in the cell and the oxygen and carbon dioxide level of whole body. In the normal body we find about 5 million erys/μl blood.

Leukocytes, or leukos, are the white blood corpuscles. (With the differential blood cell count technique the cell nuclei of the white blood corpuscles are actually blue-purple.) Leukos are very important for the immune system. Under the microscope they are clearly recognizable since they are larger than the erys. If too many leukocytes are in the blood, this can point to leukaemia (a blood cancer). The white blood cells are closely connected with the inflammation value CRP. The number of Leukos should lie between 4,4 and 11,3 x 10^8/μl*. The white blood cells can be divided into different cell types. One finds approx. 60% neutrophil granulocytes, 30% lymphocytes, 6% monocytes, 3% eosinophil granulocytes as well as 1% basophil granulocytes.

The lymphocytes are divided into B-lymphocytes and T-lymphocytes. B-lymphocytes are memory cells, which recognize pathogens and then call on the T-lymphocytes, aide cells, which destroy the infected cells.

Thrombocytes are blood platelets (not cells), they help with the closing OF wounds. If they recognize open places, i.e. wounds, they send out filaments, which blood binds too and prevents bleeding. The quantity of the blood platelets should lie between 150-360x 10^3/μl*.

* Die Normwerte sind Labor sowie auch Geräte spezifisch.

bibliography:
http://www.cholesterin -werte.com/
http://www.reventlow -apotheke.de/
http://de.wikipedia. org/wiki/Cholesterin
http://www.med4you.at/
http://www.50plus.at/
http://www.gesund heitpro.de/
http://www.marien- apotheke.or.at/
http://www.netdoktor. at/
Lit. 1 : Dr. N. Schaenzler, Laborwerte, GU Verlag