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Circulatory System The Vvertebrate circulatory system ensures the rapid transfer of bulk substances which are needed

by the body. Blood is responsible for the transportation of respiratory gases O2 and CO2, nutrients in the gastrointestinal tract, waste products, specialized cells like WBC, hormones, as well asand heat for thermal regulation (Widmaier et al, 2004). This exclusive type of circulatory system can be observed in vertebrate organisms in which vascular fluids are contained within blood vessels (Guyton & Hall, 2006). Blood flow is important because it carries particular certain substances or chemicals, which are needed by a particular organ, or a waste products which is needed tomust be secreted away from the body. Blood flows and carries materials via two routes namely: pulmonary and systemic circulations. Pulmonary circulation pertains how deoxygenated blood is being oxygenated in the lungs by passing through the pulmonary artery to the pulmonary capillaries and back to the left atrium of the heart through the pulmonary veins (Klabunde, 2012). On the other hand, systemic circulation shows how the oxygenated blood from the pulmonary capillaries is being transported into different parts of the body except to the lungs (Tortora & Derrickson, 2012). According to Klabunde (2012), there are three parts of systemic circulation: (1) Coronary Circulation where it supplies blood to the muscle of the heart, (2) Renal Circulation where 25% of the blood from the heart flows to the kidney, and lastly (3) Hepatic Portal Circulation where it supplies oxygen into the liver and carries nutrients from the hepatic portal vein.

Figure 1. Blood flow of systemic and pulmonary circulatory systems. BLOOD VESSELS Blood flow is carried out by blood vessels. In vertebrate organisms, there are three major vessels for blood transportation. Systemic circulation starts where oxygenated blood from the lungs is transported into the heart via the areteries (Barrett et al, 2012). Arteries carry out oxygenated blood from the heart to all organs of the body. Figure 3 shows the structure of a normal muscle artery.

Figure 2.

Figure 3. Structure of artery and vein. There are three major structural parts of an artery. The outer connective tissue is the tunica adventica, the middle smooth muscle is the tunica media and the inner layer connective tissue is the tunica intima (Rhoades et al, 2012). The outer and the inner laminas of the artery are responsible for the stretch and recoil mechanism during the systole and diastole processes of the heart, respectively. Arterioles as shown in figure 1 are the miniature version of asteriesarteries. They alter the degree of contraction of smooth muscle by which it allows to regulates the blood flow into different body organs (Kay, 1998). Veins have an opposite function with the arteries. It carries Oppositely, veins carry

deoxygenated blood from the different organs to the pulmonary capillaries of the lungs. Veins and arteries It hashave the same structural components along with the artery. It has alsoThe small version veins, which is thecalled venules which mediates the connection of veins and capillaries (Tortora and Derrickson, 2012). Lastly, capillaries are the smallest blood vessels in circulatory system. It connects the arterioles and venules. They are the sites for gas exchangethe exchange of gases, nutrients and other substances between the blood and the cells. Structurally, capillaries consist of thick tunica intima layer only (Barrett et al, 2012). Table 1 shows the presence or absence of structural components of the three major blood vessels. Table 1. The composition of vertebrate blood vessels. The number of + signs indicates the relative abundance of each component Component Endothelium Smooth muscle Elastic fibers Connective tissue Artery Present +++ ++++ +++ Arteriole Present ++++ ++ ++ Type of blood vessels Capillary Venule Present Present Absent + Absent + Absent +

Vein Present +++ ++ ++

FACTORS AFFECTING BLOOD FLOW Guyton and Hall, in (2006), defined blood flow as the quantity of blood that passes a given point in circulation at a given time. The overall total blood flow circulation of a normal human adult person is almost 5000 ml/min, or refereed to as the cardiac output, where it is the amount of blood pumped by the heart into the aorta. Blood flows into different kinds of blood vessels and each of these vessels has its own cross sectional area that affects the flow of fluid. Table 2 shows the characteristics of various blood vessels that affects the rate flow of blood. Vessel Aorta Small arteries Arterioles Capillaries Venules Small veins Venae cavae Cross Sectional Area (cm2) 2.5 20 40 2500 250 80 8

Base from the differences of the blood vessels, arteries and veins have the smallest cross sectional area while capillaries have almost 1000x greater area than the arteries and veins. According to (Widmaier et al, 2004), the cross sectional area of a blood vessel is inversely proportional to the rate of blood flow. As the cross sectional area increases, then there is a decrease in the rate of flow decreases. While as the rate of blood flow increases, blood pressure also increases. on the other hand, an increase in rate of blood flow, there is also in blood pressure. As a result, since in the arteries, which have small cross sectional area, then there is will be an increase in the rate of flow and decrease in blood pressure, whereas . While in the case of capillaries, there is a will be a decrease in the rate of flow while as blood

pressure increases. However, even though veins have small cross sectional value compared to the capillaries and four times greater to arteries, they blood flow do not exhibit an increasing velocity. This is due to the gravitational force that pulls down the fluid towards the center of the earth (Barrett et al, 2012). Most of the veins are located from at the lower portion of the body and carry blood upwards towards into the lungs for oxygenation and while gravity pulls it down, thus, there is a decrease in velocity and increase in blood pressure. According to (Guyton and Hall, 2006; Rhoades et al, 2013; Barrett et al, 2012), pressure is mainly observed in a non or slow flowing fluid. Table 3 and Figure 4 show the change in velocity and pressure of blood flow at given blood vessel. Cross Sectional Area 20 2500 80 Velocity Increase Decrease Decrease Blood Pressure Decrease Increase Increase

Comment [T1]: Insert reference for previous statement instead

Arteries Capillaries Veins

Figure 4 Aside from the cross sectional area of blood vessels and the gravitational pull of the earth, there are other factors that may affect the rate flow of blood namely: viscosity of the blood, chemicals that (may dilate or constrict the blood vessels), varying temperature and mechanical pressure. The same withSimilar to water, blood has its own viscosity (and it is 3-4 times as viscous as water). The viscous property of the blood is highly affected by the bloodits compositions. Blood carries various substances with varying sizes such as ions, excess leukocytes, erythrocytes and plasma, thus, the rate flow of the blood decreases. This process is referred as the Poiseuille-Hagen principle where the volume flow rate of the blood is inversely proportional to viscosity (Barrett et al, 2012). Capillary Physiology Capillaries are highly distributed all around the body. It is connected to the arteries through arterioles which are divided into smaller vessels referred as the metarterioles. The openings of the capillaries are surrounded by a ring of smooth muscle called known as the precapillary sphincter (Klabunde, 2012). This smooth muscle is responsible for the relaxation or contraction of the capillaries in response to metabolic stimulus (Kay, 1998; Widmaier et al, 2004). Even though each of the capillaries has smaller cross sectional area, but the total sum of all the cross sections sectional areas of these vessels is 3-5x greater compared to the arteries and veins (Guyton and Hall, 2006). The Ccapillary bed is the site where ions and nutrients diffuse. There are three processes where substances move across the capillary walls namely: diffusion, through vesicle transport, and bulk flow mechanism (Guyton and Hall, 2006; Widmaier et al, 2004). Capillaries contain only one layer of endothelial cells. Lipid soluble ions such as oxygen and carbon dioxide can easily diffuse through the plasma membrane of the endothelial cells. On the other hand, water filled channels which are mainly found in intracellular clefts of the capillary blood vessel which allows ions and polar molecules to diffuse across the membrane. However, these water filled channels allow only small size proteins to be transported, thus, medium sized proteins and substances are transported through vesicles. Substances

Comment [T2]: Kulang sa thought ano ang purpose ng water filled channels?? OR must be: On the other hand, water filled channels, which are mainly found in intracellular clefts of the capillary blood vessel, allows ions and polar molecules to diffuse across the membrane.

are endocytosed from the plasma of the luminal border and exocytosed at the interstitial side of the membrane (Barrett et al, 2012). Lastly, the bulk flow mechanism is not for exchange of ions and small substances but, rather, for the distribution of extracellular fluids (Barrett et al 2012; Rhoades et al, 2013). VASCULAR COLOR OF THE MEMBRANE Colors of the membrane depend on the reaction of the capillaries into different stimulus. When these blood vessels are dilated and blood flow is taking place through them, the color of the capillaries is red. If On the other hand, whenthe blood stagnates in relaxed capillaries, the oxyhaemoglobin goes into a series of reduction reactions, thus the skin capillaries is become blue. Lastly, and lastly, when the capillaries are contracted, the color of the skin isbecomes pale (Keele and Neil, 1965; Klabunde, 2012) The main objective of this study is to determine the effects of different chemicals, varying temperature and mechanical pressure on the peripheral and cardiac capillary blood flow of the anuran circulatory system. RESULTS AND DISCUSSION Vertebrate animals, particularly mammals, evolved a more advanced cardiovascular system. This mechanism which helps them organism to attain homeostasis through altering the cardiac output and changing the diameter of the different blood vessels (Barrett et al, 2012). In Aaddition to that, metabolites and other chemicals may also contribute on the adjustments of vessels to attain a stable system. These chemicals can be classified into two groups namely: vasodilators and vasoconstrictors. Vasodilation is a process in which metabolites or chemicals may result intoinduce the widening of the blood vessels through the action of vasodilators. This can be achieved when there is a decrease in diameter of the blood vessels thus, blood pressure decreases but the rate flow of the blood flow increases. On the other hand, vasoconstriction exhibits a narrowing process in of the diameter of the blood vessels. This which contradicts to the mechanism of vasodilation into different types of blood vessels (Barrett et al, 2012). Table 4 shows the effects of the different metabolites, chemicals, varying temperature and mechanical stress into the peripheral and cardiac capillaries of the anuran systemic system. Table 4. Effects of vasodilating chemicals into the peripheral and cardiac capillaries. Solutions Adrenaline Acetic acid Lactic acid Nicotine Alcohol Warm water Ice water pithing Peripheral Capillaries Decrease Decrease Decrease Decrease Increase Increase Decrease Decrease Cardiac CApillaries Increase Increase Increase Decrease Increase Increase Decrease increase

Comment [T3]: Capillaries dapat since capillaries ang pinaguusapan Comment [T4]: Red?

ADRENALINE

Result showed that adrenaline decreases the rate flow of blood flow in peripheral capillaries. This is due to the interaction of the adrenaline molecules with to the receptors of epinephrine namely, alpha-1 and beta-2 adrenergic receptors, which are responsible for the vasoconstriction and vasodilation to of the different blood vessels. Peripheral (peripheral blood vessels contain both receptors). Adrenaline molecules are more attracted to alpha-1 receptors but only for a short period of time. howeverHowever, alpha-1 adrenergic receptors is are more abundant compared to the beta-2 receptors, and thus constriction of the smooth muscle occurred (Klabunde, 2012; Guyton & Hall, 2006). In contrarycontrast, the effect of adrenaline to the heart causes caused it toan increase in blood flow, thus, there is lesser blood pressure. Again, both alpha-1 and beta-2 adrenergic receptors are present in cardiac capillaries of the heart. Cardiac capillaries exhibit biphasic response to the adrenaline molecules. Adrenal molecules first bind to the alpha-1 receptors but for a short period of time and causes the blood vessels to contract. However, the molecules leave the alpha receptor and binds to the beta-2 adrenergic receptor for a long period of time which will results into vasodilation (Barrett et al, 2012). According to Keele and Neil (1965), adrenaline raises the systolic blood pressure, decreases the diastolic blood pressure, increases the heart rate, increases the cardiac output and reduces the blood flow in peripheral arteries and veins. Figure 5 shows the action mechanism of adrenaline in the human body. LACTIC ACID & ACETIC ACID After acidic solutions such as lactic acid and acetic acid were introduced into the capillaries, there was a decrease of blood flow (vasoconstriction) and increase of blood flow (vasodilation) in peripheral and cardiac capillaries, respectively. Capillaries have only one layer of endothelial cells and ions (e.g. H+) can easily diffuse through the plasma membrane. As H+ increases (decrease pH), vascular smooth muscle regulates their own blood flow through autoregulation. Blood vessels respond to the extreme environment by contracting and thickening the surface area of the blood vessels, thus blood pressure increases. For the dilation of the cardiac capillaries, introduction of acidic solutions caused an increase in the resistance of the arteries against the blood flow. This rapid blood flow occurs to contradict the acidic environment of the cardiac capillaries. The same result was observed when Fleishman et al (1957) determined the effect of pH change upon systemic large and small vessel resistance. Acidic solutions, particularly lactic acid, produces hydrogen ions which are increased through the bicarbonate buffer system when CO2 releases. From glycolysis, pyruvic acid can undergo fermentation to form lactic acid and increases H+ which results into vasodilation. The rapid blood flow is associated with the extraction of oxygen to meet the metabolic needs of the organ tissues. This is referred to as the active hyperaemia, which results from the . This is due to increase in H+ and, CO2, and decrease in O2 concentration, thus, arterioles dilated and blood flow increases. When O2 decreases, there is limited production of ATP which is important for the contraction by ofthe cardiac muscle. Depletion of ATP results to causes the ATP molecules the inhibition of the K+ channels, which will result into prolonged hyperpolarization of the endothelial cell membrane., tTherefore, voltage the activity of voltage-gated calcium channels activity is reduced and causes vasodilation (Klabunde, 2012). NICOTINE

When nicotine decoction extract was applied into the capillaries, both peripheral and cardiac capillaries showed a decreasing blood flow rate rate flow of blood. This is due to the mechanism of nicotinic acetylcholine receptors which activates ligand- gated channels. When nicotine molecule binds to the nicotinic acetylcholine receptor at of a vascular smooth muscle cell (tunica media), it releases VEGF, FGF2 and PDGF. These three releasing factors of nicotine cause the migration and aggregation of vascular smooth muscle cells which surround the endothelial cells into the basal side of the blood vessel. This migration results into a gap or space between the smooth muscle and the endothelial cells. Macrophages would tend to accumulate at the space, thus, clogging the passage way of the blood. As a result, blood flow decreases due to high resistance of the blood to the blood vessel wall, radius of the lumen of the vessel decreases and vasoconstriction occurs ( Barrett et al, 2012; Klabunde, 2012; Rhoades & Bell, 2013).

ALCOHOL When ethanol was introduced into the peripheral and cardiac capillaries, there was an increase of in blood flow rate. This is due to the heat generated by the alcohol. The catabolic process of alcohol releases a lot of ATP (exothermic reaction) which. This release of ATP causes an increase in body temperature. This vascular effect is due to the combined effects of released histamine, 5-hydroxytryptamine and vaso-active polypeptides. The local heating causes a vasodilation in capillaries and later to the arterioles (Keele & Neil, 1965). Also, blood maintains alcohol or ethanol content (BAC). When alcohol concentration increases, blood flows rapidly to prevent the inhibitory effect of alcohol where GABA receptor is inhibited by GABAA neurotransmitter that increases the flow of Cl- in postsynaptic terminal. This accumulation of Cl- causes prolonged hyper-polarization and inhibits depolarization, thus action potential generation is inhibited (___???reference).

WARM WATER and COLD WATER When warm water was introduced into the peripheral and cardiac capillaries, results showed that the capillaries in both areas were dilated. This occurs due to the stimulation of hypothalamic neurons by the warmed blood. Thus theThe neurons of the anterior part of the hypothalamus are sensitive to the temperature of the blood supplying them and cause vasodilation. On the other hand, when cold water was applied into the capillaries of peripheral and cardiac muscles, the blood vessels were vasoconstricted. This is due to the autoregulation of the blood vessel which was. This made possible because the hypothalamic centers are both stimulated both directly by the cooled blood reaching them and through by the afferent impulses from the stimulated area. Additionally, The vasoconstriction may lead into increases in blood pressure (Klabunde, 2012). PITHING When mechanical pressure such as pithing was applied into the peripheral capillaries, a decrease in the rate of blood flowof rate flow of blood was observed, while there was an increase of blood flow in cardiac capillaries. Stimulus such as crushing or destroying the muscle causes the

capillaries to release endothelin from the endothelial cells through the action of endothelin converting enzyme (ECE) (Klabunde, 2012). In blood vessels, the release of endothelin allows the contraction of the vessel as large as 5 millimeters in diameter of the endothelial cells to prevent extensive bleeding (Guyton & Hall, 2006). Endothelin also functions for the contraction of the adjacent vascular muscle of the blood vesseles (reference). The endothelial cells release the endothelin-1 which binds to the adjacent vascular smooth muscle. The Aadjacent vascular smooth muscle contains two types of receptors namely: ETa and ETb. These two receptors are coupled by Gq protein which is responsible for the formation of IP3 (inositol triphosphate) from phosphatidylinositol (PIP) as shown in figure __. As The increase of in IP3 allows the release of Ca2+ in the sarcoplasmic reticulum which causes the contraction of the muscle (Klabunde, 2012). On the other hand, the release of endothelin causes the cardiovascular capillaries to vasodilate. Mechanical pressure such causes induces the endothelial cells to release nitric oxide, formerly known as endothelium derived releasing factor (EDRF), produced from the amino acid L-arginine by the enzymatic action of nitric oxide synthase. There are two forms of the endothelial nitric oxide synthase: constitutive nitric oxide synthase (cNOS) and inducible nitric oxide synthase (iNOS). At the basal side of the blood vessels, nitric oxide is continually produced by the constitutive nitric oxide synthase and the enzyme is usually mediated by calcium and calmodulin. There are two pathways where nitric oxide is being released. First, nitric oxide is formed by the rapid blood flow that causes shearing on the endothelial cells, thereby releasing which causes the release of Ca2+ and activates constitutive NOS. This first mechanism is also referred to as flow-dependent nitric oxide formation. The second mechanism is the receptor-stimulated nitric oxide formation. Variety of ligand receptors such as bradykinin and acetylcholine (Klabunde, 2012; Rhoades & Bell, 2013) triggers the release of Ca2+ and allows the formation of nitric oxide. The presence of nitric oxide, which only lasts for few seconds, allows induces the blood vessels to vasodilate. At intracellular level, after the formation of nitric oxide at vascular endothelial cells, it rapidly diffuses into the adjacent vascular smooth muscle and, binds to the heme moiety and activates the guanylyl cyclisecyclase. This enzyme dephosphorylates GTP into cyclic 3,5guanosine monophosphate (cGMP) (Barrett et al, 2012) which serves as the messenger for the relaxation of the smooth muscles. CONCLUSION

Blood vessels are important for the transportation of ions and nutrients in to the other parts of the body through the blood. Capillaries are small blood vessels where exchange of ions and bulk nutrients occur. This study was done to determine the effects of the different solutions in peripheral and cardiac capillaries. Results showed that adrenaline, acetic acid and lactic acid and mechanical pressure (pithing) cause decreases the rate of blood flow a decrease of blood flow in peripheral capillaries, resulting into vasoconstriction, while and increases the rate of blood flow there was an increase in rate flow of blood in cardiac capillaries, causing vasodilation . This is due to the mechanisms actions of receptors, ions and chemical substances that trigger the endothelial cells of the different blood vessels, particularly the capillaries to relax or to contract. Nicotine causes the accumulation of macrophages in

arterial blood vessels which result into congestion of blood flow and vasoconstriction in peripheral and cardiac blood vessels. While, alcohol causes an increase in blood flow in peripheral and cardiac capillaries. Lastly, cold and warm temperature causetemperature causes an vasoconstriction and vasodilation in peripheral and cardiac capillaries, respectively, to regulate rate flow of the blood.

Abstract This study was undertaken to determine the effects of the different solutions and stresses in peripheral and cardiac capillaries of anuranan Anuran. Results showed that adrenaline, acetic acid, lactic acid and mechanical stress (pithing) cause decreases in rate flow of bloodthe rate of blood flow in peripheral capillaries while and there was an increases the rate of blood flow in blood in cardiac capillaries. On the other hand, both nicotine and cold water result into vasoconstriction of the blood vessels while alcohol and warm water result into vasodilation in both peripheral and cardiac capillaries. These vasoconstriction and vasodilation of the blood vessels allow regulating the regulation of blood in the body system.