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Zinc may be complexed with calcium-phytate and lead to inefficient utilization of dietary zinc arteria labialis superior effective 8mg aceon. Adequate calcium intake will compensate for the reduced availability of calcium blood pressure monitor watch purchase aceon online from canada, but this will aggravate the zinc deficiency and such a relationship has frequently led to parakeratosis in pigs hypertension icd 9 discount 4mg aceon mastercard. Phytic acid reduces the absorption of calcium from the gastro-intestinal tract and consequently implicated in the development of rickets when chicks are fed cereals such as sorghum blood pressure medication beta blockers side effects generic 8 mg aceon. Zinc and iron deficiency symptoms have been reported in man and chicken (Lease, 1966) when fed diets high in phytic acid. There is a significant inverse relationship between phytic acid content and the availability of calcium, magnesium, phosphorus and zinc in products like soya bean, palm kernel, rapeseed and cottonseed meals (Nwokolo and Bragg, 1977). Manganese Mn 2+ Active in formation of amino acids, activates some enzymes, coenzyme activity, required for water-splitting step of photosynthesis, chlorophyll synthesis Active in formation of chlorophyll, activates some enzymes, plays a role in formation of auxin, chloroplasts, and starch 2+ Light green leaves with green major veins, leaves whiten and fall off Zinc Zn 2+ Chlorosis, mottled or bronzed leaves, abnormal roots Copper Cu,Cu + Component of many redox and lignin-biosynthetic enzymes Essential for nitrogen fixation, cofactor that functions in nitrate reduction, component of enzyme used in nitrogen metabolism Cofactor for metabolism an enzyme functioning in nitrogen Chlorosis, dead spots in leaves, stunted growth, terminal buds die, necrosis in young leaves Possible nitrogen deficiency, pale green, rolled or cupped leaves, mottling and necrosis in old leaves Molybdenum MoO4 2- Nickel Ni 2+ Source: (Williams, 1992). Sometimes, either as a conse-quence of low availability of iodine from the soil, or the presence in the food of goitrogenic substances, which interfere with the utilization of iodine by the thyroid, the iodine requirements of some animals. These relationships make it difficult to determine the optimum dietary level for the individual elements required for humans and domestic animals. As a result of this, the recommended dietary level of any element should rarely be considered independent of the level of other essential nutrients (Hays and Swenson, 1985). Examples are the definite relationship of calcium and phosphorus in the formation of bones and teeth and as the major structural elements of the skeletal tissue. Hypocalcaemia may cause weakness of the heart similar to that caused by hyperkalaemia. A high level of potassium appears to increase the requirement for sodium and vice versa (Merck, 1986). Potassium deficiency leads to an increase in the basic amino acid concentration of the tissue fluids and some increase in cellular sodium levels as a means of maintaining cation-anion balance. Potassium influences the contractibility of smooth, skeletal, and cardiac muscles and has an effect on muscular irritability that, like that of sodium, tends to antagonize the effect of the calcium ion. Under conditions of salt restriction, calcium appears highly important in helping to maintain the potassium content of tissue (Hays and Swenson, 1985). There are also inter-relationships of iron, copper and cobalt (in vitamin B12) in haemoglobin synthesis and red blood cell formation (Hays and Swenson, 1985). Copper deficiency results in an increase in iron in the liver, whereas an excesss of copper results in a decrease in iron content of the liver, thus reflecting the role of copper in iron utilization. Copper is present in blood plasma as a copper-carrying plasma protein called erythrocuprin. Erythrocuprin provides a link between copper and iron metabolism and mediates the release of iron from ferritin and haemosiderin (Hays and Swenson, 1985). The dietary requirement of copper is affected by the level of some other minerals in the diet, and is increased in ruminants by excessive molybdenum. Treatment of copper poisoning is based on the rationale that excess molybdenum may cause copper deficiency and molybdenum in conjunction with the sulfate ion has been used in treating copper poisoning in ruminants (Pierson and Aenes, 1958). The Cu requirement varies among animal species to some extent but is influenced to a large degree by its relationship with and the intake of other mineral elements such as iron, molybdenum and sulfate. Sodium, potassium, calcium, phosphorus and chlorine serve individually and collectively in the body fluids. Under stress conditions, a loss of sodium may be compensated for by an increase in potassium; but the animal is limited in its capacity to substitute bases, and major losses of sodium lead to a significant lowering of osmotic pressure, and therefore to a loss of water or dehydration. In animals, excess chloride and a constant level of sodium can result in acidosis, whereas an excess sodium and a constant level of chloride can result in alkalosis. Intake of excess dietary manganese had been reported to interfere with phosphorus retention (Hays and Swenson, 1985). The amount of copper in the diet of animals necessary to prevent a copper deficiency is influenced by the intake of other dietary constituents, notably molybdenum and inorganic sulphate (Merck, 1986). High intake of molybdenum in the presence of adequate sulphate increases the requirement for copper in animals (sheep) (Merck, 1986). Cardiac muscle, skeletal muscle and nervous tissue depend on a proper balance between calcium and magnesium ions. The symptoms of magnesium deficiency resemble that of low-calcium tetany (Hays and Swenson, 1985). Deficiencies of trace elements like zinc, copper and magnesium have been implicated in various reproductive events like infertility, pregnancy wastage, congenital anomalies, pregnancy-induced hypertension, placental abruption, premature rupture of membranes, still births and low birth weight (Pathak and Kapil, 2004).

A culture is considered a primary culture up to its first subculture or passage; after that it is named a cell line blood pressure chart stage 1 hypertension aceon 2 mg free shipping. Animal cells: basic concepts 19 shows a schematic diagram of the sequence of events associated with subculture of a normal primary cell culture arrhythmia associates fairfax va cheap aceon 2 mg amex. Primary cultures are initially heterogeneous heart attack marlie grace discount aceon uk, but fibroblasts may become predominant after some growth hypertensive emergency purchase aceon 4 mg with amex. Obtaining primary cultures is laborious and cells can be maintained in vitro for only a limited period. During this period, cells generally maintain the differentiated characteristics of the original tissue from which they were harvested. For instance, the enzymatic dissociation of organ fragments or adherent cells breaks cell­cell or cell­surface interactions. Dissociated cells generally change their shape, becoming rounded and losing their phenotypic polarity, modifying protein distribution in cell membrane. Certainly not all cells survive cell manipulation but those that survive should be able to correct any injuries and adapt to environmental changes. Cells condition their environment through the release of substances into culture medium, such as growth factors, which promote cell adherence and proliferation. Some cell lines can adapt, proliferate, and differentiate faster if culture medium is previously conditioned by cells in active growth. Environmental conditioning can also be achieved by adding cells to medium already inoculated with a non-mitotic viable cell population, named feeder cells. These cells can be prepared from 3T3 fibroblastic cells exposed to gamma radiation or treated with mitomicin C (Rheinwald, 1989). After successive subcultures of a very heterogeneous primary culture, containing many cell types of the original tissue, a more homogeneous cell line with a higher growth rate may arise. A cell line can be serially propagated in culture, usually for only a limited number of cell divisions. Finite cell lines are generally diploid and maintain some degree of differentiation. Nevertheless, these cell lines die after a limited number of generations, the Hayflick limit, which is usually about 30­50 division cycles depending on the origin of the cells (Hayflick and Moorhead, 1961). The immortalization of a cell line can be accomplished as a spontaneous process, by an oncogene or virus or by chemical treatment. This can lead to a continuous cell line that can be propagated for an undetermined period. If such changes occur with an affect on cell cycle control, the cell line can be designated a transformed cell line. According to Freshney (2000), the main characteristics of transformed cell lines are: (i) altered cell morphology (smaller, less adherent, or more rounded cells, with a higher nucleus to cytoplasm ratio); (ii) higher growth rate (duplication times decrease from 36­48 hours to 12­36 hours); (iii) less dependency on blood serum or selected growth factors; (iv) increase in cloning efficiency; (v) increase in heteroploidy (chromosomal variation between cells) and in aneuploidy (divergence from the original diploid number); (vi) increase in tumorigenicity. The main advantage of transformed cells for cell culture is the almost unlimited cell supply. However, a disadvantage is that the cells generally maintain few characteristics of their original tissue. Culture medium changes should be performed even in cultures showing no cell proliferation, since cells can metabolize and deplete nutrients from the medium. Intervals between medium changes and subcultures may vary depending on the cell line, growth rate, and metabolism. An increase in cell density, pH decrease, nutrient depletion in the medium, or alteration in cell morphology indicate the need for culture medium replacement. These cells normally proliferate in monolayers and show contact inhibition, with the maximum cell yield generally limited by the available surface of the culture vessel. Blood cells (such as those derived from lymphomas) generally grow in suspension, while cells derived from solid tissues (such as kidney and liver) are adherent cells. Since adherent cells proliferate only after cell surface adhesion, an understanding of the steps of this process is very important. The first step consists of the adsorption of adhesion factors to the substratum, such as Table 2. Animal cells: basic concepts 21 vitronectin and/or fibronectin glycoproteins and often associated to Ca2ю ions. These factors can be derived from serum or can be produced by the cells themselves.

Introduction to animal cell technology ґ Paula Marques Alves blood pressure medication with little side effects aceon 2mg sale, Manuel Jose Teixeira Carrondo blood pressure normal lying down generic aceon 2mg online, and Pedro Estilita Cruz 1 1 blood pressure yoga breathing exercises buy generic aceon from india. Nevertheless blood pressure keeps spiking cheap aceon 2 mg line, the first experience with animal cell culture can be traced back to the beginning of the 20th century. By the use of the hanging drop technique and frog heart lymph, Ross Harrison, at Yale, tried between 1906 and 1910 to elucidate how the nervous fiber is originated (Witkowski, 1979). He considered three hypotheses: (i) in situ formation from the nerve sheath; (ii) preformed protoplasmic bridges; or (iii) as a result of the nerve cell growth itself. When Harrison demonstrated the validity of the third hypothesis, he also confirmed the cell as the primary developing unit of multicellular organisms. An early pioneer of cell culture was the French surgeon Alexis Carrel, who won the Nobel prize in Medicine in 1912 for his research at the Rockefeller Institute (Spier, 2000). Harrison was, above all, the inventor of analytical solutions, while Carrel, with his extensive clinical practice experience, sterility concerns, and capacity to develop appropriate culture media and culture flasks, created the change in the technological paradigm that led to the start up of animal cell technology. By careful manipulation, Carrel insured the maintenance of chicken embryo cells for several decades in culture. Spier (2000) lists some essential differences between cells in an organism (in vivo) and cells in culture (in vitro), particularly the following; Tissues are three-dimensional, while cell cultures are of zero dimension (monodispersed in suspension culture) or two-dimensional (monolayer growth). Requirements to avoid contamination have led to the formulation of serum-free media or even of protein-free, chemically defined media for the production of biopharmaceuticals (Griffiths, 1988). Fixed- and fluidized-bed bioreactors, air-lift reactors and even stirred and aerated tanks with capacities up to 15 m3 are common in large plants producing monoclonal antibodies (mAbs) for anticancer therapies (Adams and Weiner, 2005; Griffiths, 1988). One of the main purposes of animal cell culture development was the search for viral vaccines, initiated during the Second World War (1939­ 1945), particularly for poliomyelitis. Later, after a dispute between Hilary Kaprowski and Albert Sabin, the attenuated vaccine against poliomyelitis was licensed in 1962. At the end of this period, at the Wistar Institute, Hayflick developed a cell line from embryonic tissue capable of replicating more than 50-fold before becoming senescent (Hayflick and Moorhead, 1961). The cell was diploid, easy to freeze and to reactivate and did not show any evidence of contamination by the viruses normally found in monkey primary kidney cells. Finally, three relevant aspects should be mentioned to clarify the scientific, technological, and industrial position of biopharmaceuticals and animal cells. This means that analytical, biological and immunological characterization assays are usually not considered sufficient for product marketing, given the complexity of the molecules. Therefore, product licensing is based on the specific production process, which cannot be altered. Process changes may require new licensing proce- Introduction to animal cell technology 3 dures, and this makes the introduction of biosimilars in the market more difficult. In the last few years, market dominance in favor of animal cells has increased significantly. This may be different from what is normal in the production of simple biological compounds such as antibiotics or vitamins. The number of biological processes has increased tremendously in the last 15 years. This has resulted in high expectation for an improvement of quality of life and an increase in the volume of business related to products obtained from animal cell culture technology, with the broad potential use of these products for disease diagnostics, prevention, therapy, and cure. However, the huge growth and expansion of this technology was possible only because new cell types were established, namely diploid cells, hybridomas, and other continuous cell lines. However, depending on their applications, animal cells can also be grouped as follows. Primary cells are normally heterogeneous and better represent the tissue from which they originate. These cells have a finite growth capacity and can be subcultured for only a limited number of passages. Subcultured cells, which have been selected to form a population of cells of a single type, are designated cell lines, and can be finite or continuous.

For the same reasons arrhythmia joint pain purchase cheap aceon on-line, plants pigment molecules absorb only light in the wavelength range of 700 nm to 400 nm; plant physiologists refer to this range for plants as photosynthetically active radiation heart attack while running discount 4mg aceon with amex. The visible light seen by humans as white light actually exists in a rainbow of colors hypertension values cheap aceon 2mg on-line. The visible light portion of the electromagnetic spectrum shows the rainbow of colors arteria in english buy aceon 2 mg with amex, with violet and blue having shorter wavelengths, and therefore higher energy. Violet has the shortest wavelength and therefore carries the most energy, whereas red has the longest wavelength and carries the least amount of energy. There are five major chlorophylls: a, b, c and d and a related molecule found in prokaryotes called bacteriochlorophyll. Chlorophyll a and chlorophyll b are found in higher plant chloroplasts and will be the focus of the following discussion. Each type of pigment can be identified by the specific pattern of wavelengths it absorbs from visible light, which is the absorption spectrum. Notice how each pigment has a distinct set of peaks and troughs, revealing a highly specific pattern of absorption. Carotenoids absorb in the short-wavelength blue region, and reflect the longer yellow, red, and orange wavelengths. Chlorophyll a and b, which are identical except for the part indicated in the red box, are responsible for the green color of leaves. Many photosynthetic organisms have a mixture of pigments; using them, the organism can absorb energy from a wider range of wavelengths. Some organisms grow underwater where light intensity and quality decrease and change with depth. Plants on the rainforest floor must be able to absorb any bit of light that comes through, because the taller trees absorb most of the sunlight and scatter the remaining solar radiation (Figure 8. Additional methods for the identification of plant pigments include various types of chromatography that separate the pigments by their relative affinities to solid and mobile phases. This chemical energy supports the light-independent reactions and fuels the assembly of sugar molecules. Pigments in the lightharvesting complex pass light energy to two special chlorophyll a molecules in the reaction center. The light excites an electron from the chlorophyll a pair, which passes to the primary electron acceptor. The two complexes differ on the basis of what they oxidize (that is, the source of the low-energy electron supply) and what they reduce (the place to which they deliver their energized electrons). Each photosystem is serviced by the lightharvesting complex, which passes energy from sunlight to the reaction center; it consists of multiple antenna proteins that contain a mixture of 300­400 chlorophyll a and b molecules as well as other pigments like carotenoids. The energy is transferred from chlorophyll to chlorophyll until eventually (after about a millionth of a second), it is delivered to the reaction center. Up to this point, only energy has been transferred between molecules, not electrons. What is the initial source of electrons for the chloroplast electron transport chain? It is at this step in the reaction center, this step in photosynthesis, that light energy is converted into an excited electron. The ions build up energy because of diffusion and because they all have the same electrical charge, repelling each other. To release this energy, hydrogen ions will rush through any opening, similar to water jetting through a hole in a dam. Another term, the Calvin cycle, is named for the man who discovered it, and because these reactions 232 Chapter 8 Photosynthesis function as a cycle. Others call it the Calvin-Benson cycle to include the name of another scientist involved in its discovery. However, the term dark reaction can be misleading because it implies incorrectly that the reaction only occurs at night or is independent of light, which is why most scientists and instructors no longer use it.

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