physiology

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 6:44 am

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physiology 47

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 7:09 am

نبدا بعينات w b Cs عملى.....؟

Leukocytes

The five major categories of leukocytes in mammals are neutrophils, eosinophils, basophils, lymphocytes, and monocytes. The first three collectively are called granulocytes, because they contain granules in their cytoplasm, and the latter two collectively are called mononuclear cells in reference to their round nuclei. These collective terms are somewhat misleading since some granulocytes have barely perceptible granules and most monocytes do not have round nuclei. Granulocytes of most mammals are classed as neutrophil, eosinophil, and basophil based on the reaction of their specific granules with the dyes in polychrome blood stains, called Romanowsky stains.

The prototype species for granulocyte classification is human, in which neutrophil granules are small and pink with weak affinity for azure complexes, basophil granules are deep purple because of strong affinity for the azure complexes, and eosinophils are intensely orange because of strong binding of eosin. Classification of granulocytes in animal species is based in part on homology of appearance to human granulocytes and in part on demonstration of homology of enzyme content and function.

There are many differences, some subtle and some obvious, among species and within species. This portion of the module presents the identifying features of leukocytes in several animal species and illustrates the species differences. Continue by working through the following topics

1)

(polymorphonuclear leukocytes, PMNs, polys)

The neutrophil, or the heterophil in birds, reptiles and some mammals, is the predominant granulocyte. In many species it is the predominant leukocyte in health. In health, only mature neutrophils are released from marrow to circulate in blood. Since mature neutrophils in prototypical human blood have nuclei that are segmented into lobes of condensed chromatin connected by filaments, mature neutrophils are called segmented neutrophils, or segs. The mature neutrophils of many animal species do not have a distinct physiology Neutcomp

lobe and filament arrangement, nevertheless, they also are called segmented neutrophils. The primary criterion for classification of neutrophils as either segmented (mature) or non- segmented (younger stages including band neutrophils, metamyelocytes, and myelocytes) is the shape of the nucleus. A neutrophil should be counted in the segmented category if the nucleus has regions that are distinctly constricted or has lateral protrusions resulting in irregular nuclear margins. A cell whose nuclear margins are smooth and parallel (or nearly parallel) should be counted as a band neutrophil. The presence of non-segmented neutrophils in blood is called a left shift. Inflammation is the most common cause of a left shift, but some other conditions, such as hemolytic anemia and primary diseases of marrow, also can cause release of immature neutrophils from marrow.

Normal mature neutrophils of several species are shown at the right. Canine neutrophils have white cytoplasm that contains small pink granules. Feline neutrophils have cytoplasm that is white and lacks visible granules. Equine neutrophils have white or slightly pink cytoplasm with no visible granules. The nuclei of equine neutrophils typically are long, thin and "knobby" with clumps of condensed chromatin projecting from the sides. Ruminant neutrophils have white cytoplasm with small pink granules; these impart an overall pink tint compared to the other species.

Rabbits, birds, amphibians and reptiles have heterophils. In contrast to the inconspicuous granules of neutrophils, granules in heterophils are large and stained deep orange to red. A typical avian heterophil is shown in "Blood Cells by Species".

A common and important morphologic abnormality of neutrophils is so-called "toxic change

2)
Lymphocytes

Most of the lymphocytes that circulate in healthy dogs, cats, and horses are small (mature) cells that have round nuclei with smooth, dense chromatin and a small rim of physiology Lympcomp

blue cytoplasm. These are sometimes confused with nucleated RBCs. Some lymphocytes, as illustrated by the equine lymphocyte, are slightly larger and have a finer, looser chromatin pattern. The lymphocytes found in blood of normal ruminants are quite variable in appearance. Many are large lymphocytes, as shown in the right middle panel, with loosely clumped chromatin and fairly abundant cytoplasm.

So-called "reactive" lymphocytes are larger cells with coarse (mature) chromatin, and deep blue cytoplasm as shown in the bottom left panel. Lymphocytes with these characteristics are sometimes called immunocytes and are associated with an immune response. These are fairly common in blood of young animals, especially ones that have been recently vaccinated.

Another type of lymphocyte that is found in low number in healthy animals is the granular lymphocyte, shown in the bottom right panel. Granular lymphocytes are distinguished by the presence of small pink granules collected into one area of the cytoplasm. Not all lymphocytes in blood smears are round. Some are distorted into other shapes by the mechanical forces applied to them during smearing and some are molded by contact with red cells.

3)
mono cytes.

Monocytes are extremely variable in appearance but there are no consistent species differences. Each of the four monocytes shown here as examples are from different species physiology Monocomp

but all could be found in blood of any individual animal of any species. For example, the canine monocyte shown here would feel equally at home in a smear of horse blood and the cat monocyte could have been in dog blood.

Some monocytes resemble band or segmented neutrophils, as illustrated by the canine moncyte, while others, like this equine monocyte, may be mistaken for lymphocytes. In general, monocytes are recognized as cells that have abundant light to deep blue cytoplasm and nuclei that are not round. Chromatin is reticular or only slightly condensed. Most are larger than neighboring neutrophils and lymphocytes. Small pink granules are visible in some monocytes and many monocytes have several small discrete vacuoles. Nuclear shape amongst monocytes ranges from ameboid to bean-shaped to band-like to almost segmented. The texture of cytoplasm in a monocyte is slightly grainy or coarse compared to lymphocyte cytoplasm, which is very smooth and glassy.

4)
Eosinophils

Eosinophil granules in most animals are orange but there are always exceptions. The granules in eosinophils of iguanas and some birds are actually pale blue. In general, nuclei of mature eosinophils are shorter and less segmented than neutrophil nuclei and the cytoplasm, if visible, is pale blue. Marked species variation exists regarding the number, size, and shape of eosinophil granules.

The most intraspecies variation is found in dog eosinophils. Marked variation in granule size, number and shape occurs within and between individuals of any breed. A specific and predictable morphologic variant of the eosinophil is seen in greyhounds and other sighthounds (eg, whippets, deerhounds). Eosinophils in these breeds lack visible granules and appear as cells with slightly segmented nuclei, gray cytoplasm, and vacuoles. They are sometimes mistaken for toxic neutrophils or monocytes.

Eosinophils of the cat have small rod-shaped orange granules that fill the cytoplasm, while those of the horse have very large globular orange granules. Ruminant eosinophils have many small very round orange granules.

5)
Basophils

Prototypical basophils of human blood are filled with small, round, dark purple granules. Similar basophils are found in some species but not in others.

physiology Basocomp

Canine basophils are difficult to recognize since many do not have readily apparent granules. The major identifying features of canine basophils are the long and folded nucleus, described as ribbon-like, and the unusual gray to lavender hue of the cytoplasm. Some basophils contain a few purple granules. Basophils are rare in blood of healthy dogs.

Feline basophils are packed with small, slightly oval granules that are pale lavender rather than deep purple. The nucleus in many appears to have vacuoles, which are actually granules lying on top of the chromatin. Basophils are rare in blood of healthy cats.

Basophils of horses and ruminants are similar to each other and to human basophils. These cells contain many small deep purple granules that obscure the nucleus in many cells. Arrows point to nuclear lobes in the picture of a cow basophil. Some basophils have few granules, which probably is the result of degranulation in the sample. Low numbers of basophils are commonly found in blood of healthy cattle and horses.

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 7:13 am

مُعدل تَثَفل (ترسُب) الكُرَيَّاتِ الحُمْر

مُعدل تَثَفل (ترسُب) الكُرَيَّاتِ الحُمْر Erythrocyte Sedimentation Rate, المعروف بين عامة الناس بسرعة أو بنسبة ترسيب الدم, و غالباً ما يرى الشخص الإختصار ESR على ورقة تحليل المُختبر, الذي في الغالب يُصاحب الإختصار CBC الذي هو إختصار لتحليل تعداد كامل لعناصرِ الدم Complete Blood Count الذي يتضمن عدد كريات الدم الحُمر RBC , عدد كريات الدم البيضاء WBC , عدد صفائح الدم Platelet , مستوى الهيموغلوبين HGB , مُتوسط حجم الكُرية (الحمراء) MCV , مُتوسط هيموغلوبين الكُرية (الحمراء) MCH و قياسات أخرى. تحليل سرعة ترسيب الدم تحليل غير نوعي Non-Specific Test يعني بحد ذاته لا يدُل على و لا يُشخص أي مرض مُعين أو حالة مرضية مُحددة, و لكنه دلالة عامة على وجود إلتهاب Inflammation في الجسم, سواءً كان حاداً أو مُزمناً.
تعريفه و وحدة قياسه: هو مُعدل تَثَفلِ (ترسُب) الكُرَيَّاتِ الحُمْر في فترة زمنية مُحددة (عادة الفترة الزمنية هي ساعة واحدة), و هو عبارة عن طول سقوط رأس العمود من الدم بالمليمتر في أنبوبة مخبرية, عليه تكون وحدة قياسه هي مليمتر\ساعة mm/h.
كيفية قياسه: يتم قياس مُعدل تَثَفل (ترسُب) الكُرَيَّاتِ الحُمْر ESR بطريقة ويستيرجين Westergen Method و ذلك لبساطتها و عدم الحاجة لأي مصدر طاقة, حيث تُستخدم أنبوبة ويستيرجين Westergen Tube و هي عبارة عن أنبوبة مستقيمة طولها 30 سم و قطرها الداخلي 2.55 مليمتر, و هي مُدرجة من 0-200 مليمتر و تسع حوالي 1 مليليتر.
1- يتُم تخفيف 2 مليليتر دم مع 0.5 مليليتر من 3.8%سيترات الصوديوم Sodium Citrate أو 0.5 مليليتر من 0.85% كلوريد الصوديوم Sodium Chloride.
2- يتُم ملء أنبوبة ويسترجين لعلامة الصفر (يكون الصفر في الأعلى) و تترك واقفة عمودياً تماماً في رفرف بدرجة حرارة الغرفة و بدون إهتزاز أو التعرض المُباشر لأشعة الشمس.
3- بعد 60 دقيقة (ساعة) تُقرأ المسافة من علامة الصفر إلى قمة عمود الكريات الحُمر بالمليمتر, و هذه القراءة هي قيمة مُعدل ترسُب الدم ESR.

توجد أجهزة آلية لقياس سرعة ترسب الدم مثل فس ماتك VES-MATIC 20 الذي يقيس المُعدل لعشرين عينة دم دفعة واحدة و يستغرق وقت أقل من طريقة ويستيرجين.

أي عامل في الدم يجعل كريات الدم الحُمر تلتصق ببعضها البعض و تكوين كوم تُسمى نضائد Rouleaux (كما هو مُبين في الصورة), يؤدي إلى زيادة سرعة ترسب الكريات الحُمر و بالتالي إرتفاع قيمة مُعدل ترسب الدم, و أهمها الفيبرونوجين Fibrinogen الذي يُقلل من الشُحنات الكهربية السالبة Negative Charges على سطح الكُريات الحُمر (الجهد الكهربائي زيتا Zeta Potential الذي يُبقيها مُنفصلة عن بعضها البعض في الدم), ومنها الأضداد المنّاعية Immunoglobulins و الكوليستيرول و الحمل Pregnancy و حالات فقر الدم Anaemias كذلك. و في الحالات التي يكون فيها شكل الكُريات الحُمر غير طبيعي مثل المنجلية Sickle Cells أو الكُروية Spherocytes يقل مُعدل ترسب الدم.

القيّم الطبيعية: تزيد تدريجياً مع تقدم العمر و تكون أعلى في الإناث منها في الذكور. يُمكن أن نقول بأن الطبيعي للرجال هو 1-5 و للنساء هو 5-15.

أهميته: كما ذكرنا سابقاً بأن مُعدل ترسب الدم بحد ذاته لا يدُل على حالة مرضية مُعينة, و لكنه مؤشر لوجود إلتهابات في الجسم حيث يكون مُرتفعاً في الأمراض المُعدية Infections و الأمراض المنيعة للذات Autoimmune Diseases مثل إلتهاب المفاصل الروماتويدي Rheumatoid Arthritis و السرطان, و يكون مُرتفعاً فوق 100 في مرض ألم العضلات الروماتزمي Polymyalgia Rheumatica و إلتهاب الشريان الصدغي Temporal Arteritis مما يُساعد الطبيب في التشخيص. كذلك يستخدم الأطباء مُعدل ترسب الدم في مُتابعة إستجابة المريض للعلاج, حيث أن نزوله مع العلاج يدُل على الإستجابة و عدم نزوله يدُل على عدم إستجابة المريض للعلاج. أضرب مثال, لو اشتكى شخص من آلام في المفاصل أو العضلات و تحليل سرعة ترسب الدم كان طبيعياً, عادة لا يقوم الطبيب بعمل تحاليل دم أخرى, و لكن إذا كان مُرتفعاً سيقوم الطبيب بعمل تحاليل أخرى لتشخيص الحالة.

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 8:30 am

Packed cell volume diagram

Laboratory procedure
All animals were anesthetized using 5% sodium pentobarbital, in a 25 mg/kg dose. An incision on the abdominal skin was performed to expose the abdominal cavity. Blood was collected from the abdominal aorta artery by punction. After this procedure, animals were sacrificed by the sectioning of this artery.
For hematological analyses, 1 ml of blood was collected in a test tube with an anti-coagulant substance (EDTA). A COULTER T890® was used for white cell count, red cell count, hemoglobin concentration (Hb), hematocrit (Hct), mean corpuscular volume (VCM), mean corpuscular hemoglobin (HCM), mean corpuscular hemoglobin concentration (CHCM) and platelet count. In leukocyte differential count, blood smears, fixed and stained by the Romanowisk method were used. All these procedures were performed in the Seçمo de Hematologia pf the Instituto Adolfo Lutz (Sمo Paulo - SP).
For biochemical analyses, 5 ml of blood were collected, without anti-coagulant substances, in order to obtain serum samples. The following biochemical essays were performed: total protein, serum albumin, creatinin, alanin aminotransferase (GPT/ALT) and aspartate aminotransferase (GOT/AST), using WEINER LAB kits; alkaline phosphatase and urea, using LABTEST kits. Spectrophotometric reading was performed in a continuous flow system, in a BIO 2000 (BIOPLUS®) analyzer, in the laboratory of Diagnَstico Toxicolَgico of the Departamento de Patologia of the Faculdade de Medicina Veterinلria e Zootecnia da Universidade de Sمo Paulo.
Statistical analysis
The variance analysis (ANOVA) was used to identify possible differences between hematological and biochemical values studied. P level was 0.05.

RESULTS
Values obtained for hemogram elements and for biochemical parameters from animals submitted to MEV were compared to those from the animals kept under GDV, which is the most common system used in animals facilities.
In Trial 1, significative differences were found for the following biochemical tests: albumin and urea in groups GDV and FV1 and ALT activity in the comparison between GDV and FV2 in male rats; for female rats, alkaline phosphatase values for FV1 and FV2 and urea, for the comparison GDV and FV1 and FV2 (Tab. 1).

Table 1

Values (mean ± standard deviation) for biochemical tests in male and female rats kept under Microenvironmental Ventilation System (MEV), using different levels of air speed (FV1 and FV2) and under General Diluting Ventilation system (GDV); n = 10. Sمo Paulo, 1998.

ALT = alanine aminotransferase; AST = aspartate aminotransferase; FV1: air speed level ranging from 0.03 to 0.26 m/s; FV2: air speed level ranging from 0.27 to 0.80 m/s. Different letters indicate that there are significative differences according to the ANOVA (p < 0.05)

In hemogram evaluation (Tab. 2) of male rats used in different groups in Trial 1, there were significative differences for the following components in groups FV1 and FV2: erythrocyte count, hemoglobin concentration, leukocyte count and absolute neutrophil count.

Table 2

Hematological values (mean ± standard deviation) obtained for male rats kept under Microenvironmental Ventilation system (MEV) using different levels of air speed (FV1 and FV2) and under General Diluting Ventilation system (GDV); n = 10. Sمo Paulo, 1998.

RBC = red blood cell; PCV = packed cell volume; MCV = mean corpuscular volume; MCH = mean corpuscular hemoglobin; MCHC = mean corpuscular hemoglobin concentration; WBC = white blood cell; FV1: air speed level ranging from 0.03 to 0.26 m/s; FV2: air speed level ranging from 0.27 to 0.80 m/s. Different letters indicate that there are significative differences according to the ANOVA (p < 0.05).

In the evaluation of the hemogram of female rats used in the different groups of Trial 1, there were significative differences between the number of erythrocytes of animals in groups GDV and FV2, and FV1 and FV2. In the analyses of hemoglobin values, there were differences between FV1 and FV2. In relation to hematocrit, significative differences occurred in groups GDV and FV2, and in groups FV1 and FV2. In CHCM, differences were found between GDV and FV1, and between FV1 and FV2. There was also a significative variation in the comparison between GDV and FV1 for the percentage of eosinophils (Tab. 3).

DISCUSSION
Reference values determined for biochemical tests and hemogram elements, as discussed and emphasized in specialized literature, may not represent precisely those of a certain population or animal species and should, therefore, be carefully interpreted^4,13, once there is a wide range of physiological variation. Besides, these variations are influenced by environmental conditions, gender, age, origin, breeding system, feeding and lineage, which also may interfere with the results^9,17,19 obtained in these tests. Therefore, the most adequate procedure would be to establish laboratory evaluation reference values for every animal facility.
Results obtained in biochemical analyses in Trial 1 male groups (Tab. 1) presented significative differences, in serum albumin, urea and ALT determinations. Among the MEV female groups, there were significative differences in relation to alkaline phosphatase and urea. Although these statistically significative differences have been noted, when these results are compared to the information found in specialized reports, they are considered to be inside the normal range of variation for the animal species and lineage^4,13,16a.There is no biological reason for the variation to be clearly attributed to the management systems used in Trial 1. Alkaline phosphatase values observed in this trial were lower than the ones determined by Ringler; Dabich¹⁹. However, they were clearly higher for the male groups than for female ones, as stated by these authors.
When hematological values in Trial 1 are considered, there were significative differences between male FV1 and FV2 groups (Tab. 2), in relation to erythrocyte counts and hemoglobin concentration. When leukogram results are considered (Tab. 2), there were significative differences in the number of leukocytes and neutrophils, for both air speed levels in MEV, but not for the MEV and the GDV groups. In relation to female groups (Tab. 3), there were differences in the results for erythrocyte counts and hematocrit (FC2 X GDV; FV2 X FV1); hemoglobin values (FV1 X FV2) and CHCM (GDV X FV1; FV1 X FV2). There also was a significative difference for the relative number of eosinophils in animals of the GDV and FV1 (Tab. 3) groups. In spite of these evidence, values found were inside the range of physiological variation reported in the specialized literature^9,12,20. No clear relation could be drawn from the differences found between the ventilation systems used (MEV and GDV).
When biochemical evaluation of the animals in Trial 2 is considered (Tab. 4), only alkaline phosphatase values for MEV groups IT7 and IT9 were significatively different from those of the GDV group. As already stated, these values were lower than the ones found by Ringler; Dabich¹⁹. However, this finding may be connected to physiological lineage variation, and environmental conditions, among other factors, for the values for alkaline phosphatase obtained were similar to those found by other authors^4,13,16.
In relation to the variation of erythrogram elements (Tab. 5), in Trial 2 there were significative differences for the number of erythrocytes (IT3 X IT9), for HCM ((GDV X IT3; IT3 X IT9) and for platelet counts (GDV X IT9 and IT5 X IT9). However, values found in literature also place these results inside the normal range of variation^5,19. Leukogram evaluation (Tab. 5) showed significative differences for the number of leukocytes and lymphocytes, when results for the GDV group and MEV groups with the smaller bed change interval (IT3 and IT5) were compared. These groups presented the lowest values for these counts. The similarity between the results for these counts in GDV animals and MEV groups with larger intervals for bed change (IT7 and IT9) may suggest a lower stress level in a more comfortable situation. It should be emphasized that IT3 and IT5 results were inside the normal range of variation for the species found in specialized reports^4*.
Using the same animals of Trial 2, Carissimi⁶ studied ponderal development and feed intake of animals kept under MEV and GDV, and demonstrated that there was no significative difference for ponderal development in animals kept in both systems. In relation to feed intake, rats submitted to MEV ingested a statistically higher quantity than the ones submitted to GDV, no matter the bed change interval used. However, this fact did not produce any differences in erythrogram or total protein and albumin variations, in the comparison of the results for both groups. Therefore, intake observed may be related to higher energy consumption for body temperature maintenance in animals submitted to MEV.
It was noted that statistical differences observed occurred in a disperse way, and none of the systems studied was favored. Each of the items analyzed seem to point out for one of the ventilation systems, when they are analyzed individually. However, when all the items of each biochemical / hematological parameter are evaluated as a group, no correlation is found. It may be concluded that the differences observed are result of casual variability.

CONCLUSION
Results obtained in the present research trial demonstrate that the use of the MEV system does not produce significative alterations on biochemical or hematological parameters studied. This fact is significative and extremely important, for researchers should always be concerned with the standardization of laboratory animals, because they will influence one of the basic principles in research: result reproducibility. The need for determining reference values for different laboratory exams in every animal colony should be emphasized, because of the different and countless influence factors to which they are submitted in animal facilities. Reproductive, sanitary and bed change interval advantages of the MEV system, besides the data obtained in this trial and the possibility of higher control of microenvironmental variables suggest that this ventilation system should be used in the atmospheric control of animal facilities.

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 8:38 am

Intended Use:

This fast, easy to use blood typing kit will give your clients or patients valuable information using only a finger-stick. The Eldoncard is a patented test card device that comes specially treated and impregnated with dried antibody sera Anti-A, Anti-B and Anti-Rh used universally for determining Blood Type by the forward typing method. After only a minute of coagulation, the test card to the left can be compared to the Results Chart (see below) to determine the correct blood grouping.

Background:
Blood typing is determined by the type of antigens or markers that are on the surface of red blood cells (either "A" or "B") and if there are antibodies to a portion of the blood type known as the Rh factor (either "positive" or "negative"). Blood type is inherited from, and dependent upon, the blood type of an individual's parents. In the case of a transfusion, a person's blood type needs to be compatible with the donor's blood type or an allergic-type reaction can occur resulting from the immune system of the recipient attacking the incompatible donor blood cells as invading organisms. Also, it is also very important to know the Rh factor status when woman is pregnant. If a pregnant woman is Rh negative and her baby is Rh positive, the mother's immune system can sometimes attack the baby's blood cells because of the differing blood types. All pregnant women should routinely have a blood typing test done to see if they are at risk. This blood typing should generally be done as part of routine prenatal care.

Interpretation of Results:

Included with each test kit is a color results chart shown below. This chart illustrates the different test results possible on the Eldoncard. After the blood typing test is completed and fully developed, the Eldoncard will resemble one of the graphics on the chart. The blood type and Rh factor can now be determined by matching the results on the card with the chart below.
physiology Bloodchart

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 10:12 am

Blood Coagulation

الموضوع بالشرح كامل على الرابط:-

http://www.thrombin.com/BloodCoagulation.phtml

من طرف **dr:sniper** الأربعاء 16 أبريل 2008, 10:14 am

A basic overview of the physiological mechanisms of normal hemostasis.	Tests used to diagnose disorders of hemostasis in domestic animals.
Specific information on the signs and mechanisms of disorders of hemostasis affecting domestic animals.	Knowledge of blood groups and transfusion products is essential for appropriate treatment of hemostatic disorders.

physiology

physiology

رد: physiology

رد: physiology

رد: physiology

Blood groups

Blood Coagulation

رد: physiology