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The size of the packed bed is a critical parameter heart attack 50 years buy betapace online pills, since the dissolved oxygen concentration in the medium Bioreactors for animal cells 231 gradually decreases as the liquid flows through the bed (Fassnacht and Porter blood pressure chart conversion discount 40mg betapace amex, 1999) blood pressure chart infants purchase betapace 40 mg otc. One of the advantages of this system is that a clarified fluid is obtained at the bioreactor outlet, with no need for a cell separation device. Fluidized-bed bioreactors Fluidized-bed bioreactors are also used mainly for the culture of cells attached to microcarriers. In these bioreactors, a given amount of microcarriers is fluidized inside a compartment of the bioreactor, avoiding cell damage due to bubble explosion. These bioreactors are operated in perfusion mode and cell concentrations reach 5 to 10 million cells per mL of medium. Several authors report their results in terms of cell concentration per microcarrier volume and, in this case, values in the order of 20 to 35 million cells per mL of carrier are obtained (Wang et al. Bioreactors providing surfaces for attached cell growth Roller bottles 1 systems with multiple plates, such as Cell Factory (Nunc) and and CellCube (Corning), provide a large surface area for cell adhesion and proliferation. In the case of roller bottles, although the relative surface area (area/volume) is small and consequently the production capacity is low, large-scale plants can be established by using a large number of roller bottles. This approach was adopted by the company Amgen for the industrial production of erythropoietin. Operation costs for this type of plant are high, because they are labor-intensive, and the risk of contamination is high, due to the manipulation of lots of small bottles. The latter consists of multiple trays, providing surface areas in the range of 70025 000 cm2, for1 medium volumes of 0. The system called CellCube provides a surface area in the range of 850085 000 cm2 and is fitted with an automated control system (Corning, 2006). Hollow-fiber bioreactors the core part of a hollow-fiber bioreactor is the hollow-fiber membrane module, also simply known as the cartridge. It consists of a plastic cylinder containing hundreds of semi-permeable capillary tubes, known as hollow fibers. The fibers can be made of different materials, such as cellulose esters and polysulfone. The pore size of fibers commonly employed in animal cell culture corresponds to a molar mass cut-off between 10 and 100 kDa. Low-molar-mass molecules, such as glucose and ammonia, can move freely through the pores of the fibers, at a rate that is controlled just by the pressure gradients generated by the medium recirculation pump. Inhibitory metabolic products, generated as a consequence of cell growth, diffuse in the opposite direction and get diluted in the recirculating culture medium. In the same way, low-molar-mass proteins and growth factors produced by the cells or provided by serum can permeate the fibers. Thus, in the axial direction of the fibers a decreasing gradient of medium components and an increasing gradient of metabolites are formed. This process may lead to a decrease in the transfer of nutrients to the microenvironment of the cells, with the possibility of the concentration of these molecules becoming growthlimiting. This may become critical when fiber length is increased, since a region very poor in nutrients and very rich in inhibitory metabolites can be created in the final portion of the fibers. Culture medium containing serum can be added to this recirculation stream, while basal medium is recirculated through the lumen of the fibers. Bioreactors for animal cells 233 Intracapillary circuit Extracapillary circuit Pressure sensor Pressure sensor Hollowfiber cartridge Extracapillary chamber Intracapillary chamber Recirculation pump Basal medium Residue Oxygenator pH sensor Product harvesting Serum-containing medium Figure 9. Oxygen transfer is an important issue in the operation of a hollow-fiber bioreactor. In these bioreactors, a membrane-based aeration system is usually included in the intracapillary recirculation loop to enrich the recirculating culture medium with oxygen. However, due to the low solubility of oxygen in aqueous solutions, the recirculation speed through the fibers must be very high. It is possible to modify some design parameters to determine optimal operational conditions for these bioreactors, such as the fiber material, the packing density of fibers inside the module, the dimensions of the module, the pore size of the fibers, and the type of aeration system used in the recirculation loop. However, these parameters are highly dependent on variations in nutrient feed and metabolic products removal (Gramer et al. On the other hand, an increase in product titer and an improved metabolic 234 Animal Cell Technology efficiency in terms of product formation can be attained if an operation strategy is adopted that includes a perpendicular extracapillary recirculation stream and a regular purge of cells, to operate at lower biomass concentrations (Rodriguez et al. The optimized strategy provided a significant decrease in the ratio of the flow rates of product harvest and medium feed, with a twofold increase in the total amount of antibody harvested and a sixfold decrease in the harvested volume. Thus, antibody concentration in the product stream was increased by over 10-fold, with a positive impact on the purification process of this mAb. Although many different classifications can be adopted, the most general is the one that considers the following operation modes: batch, fed-batch, continuous, and perfusion, which is a continuous mode with cell recycle/retention (Castilho and Medronho, 2002). Bioreactors for animal cells 235 kinetic expressions, such as the Monod equation, which describe the effects of nutrient and metabolite concentrations on the specific reaction rates. In order to make a decision on the most appropriate operation mode for a bioreactor, several factors must be taken into consideration. At industrial scale, the most important factors are: Characteristics of the cell line used, such as the stability of expression of the product, production pattern, degree of resistance to inhibitory metabolites, and resistance to shear stress; (ii) the market demand for the product, which determines the production scale; (iii) the technical experience of the teams responsible for both process development and regulatory issues. While substrates are metabolized, the cell population grows, forming the product and other metabolites. Figure 14B shows the concentration of cell, product (mAb), and substrates (glucose and glutamine) for a myeloid transfectoma in batch cultivation. In a batch cultivation, cell concentration increases from the inoculation density (0. Product concentration (mg/L), glucose and glutamine (mM) Cell concentration (cells/mL) 5. The range of specific product formation rates is wider, varying from a value as high as 30 pg cell1 day1, reported for mAb production by transfectomas (Velez et al. The concentration of a secreted product in batch cultures is usually in the range of 150 mg L1. After attaining maximum product concentration, the culture is interrupted and the spent medium is sent to the downstream processing stages. Due to the low solubility of oxygen, this gas must be supplied continuously, instead of just at the start of the culture, as in the case of the other nutrients. Bioreactors for animal cells 237 the batch operation mode is the simplest to carry out, and therefore it is widely employed. Generally, it is adopted for the cultivation of cells in stationary flasks, in stirred flasks, in small- and intermediate-scale bioreactors for inoculum propagation and also in several industries at production scale. In the case of inoculum propagation, the culture is stopped when the cells are still in the exponential growth phase, to provide a high mass of cells growing at maximum rate. In this way, it is guaranteed that these cells, when inoculated in a larger bioreactor, will present a minimal lag phase, and so reducing the non-productive period. The low volumetric productivities that characterize batch cultivation processes are a disadvantage for the use of this operation mode for production.
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As an example of a discrete-state machine we might consider a wheel which clicks round through 120 once a second young squage heart attack generic betapace 40 mg on-line, but may be stopped by a]ever which can be operated from outside; in addition a lamp is to arteria coronaria discount 40mg betapace light in one of the positions of the wheel fetal arrhythmia 32 weeks purchase betapace 40 mg amex. The internal state of the machine (which is described by the position of the wheel) may be q1, q2 or q3. The system of the "universe as a whole" is such that quite small errors in the initial conditions can have an overwhelming effect at a later time. It is an essential property of the mechanical systems which we have called "discrete-state machines" that this phenomenon does not occur. Even when we consider the actual physical machines instead of the idealised machines, reasonably accurate knowledge of the state at one moment yields reasonably accurate knowledge any number of steps later. As we have mentioned, digital computers fall within the class of discrete-state machines. But the number of states of which such a machine is capable is usually enormously large. For instance, the number for the machine now working at Manchester is about 2 165,000, i. Compare this with our example of the clicking wheel described above, which had three states. The computer includes a store corresponding to the paper used by a human computer. Suppose the computer is allowed 100 sheets of paper each containing 50 lines each with room for 30 digits. The logarithm to the base two of the number of states is usually called the "storage capacity" of the machine. Thus the Manchester machine has a storage capacity of about 165,000 and the wheel machine of our example about 1. This leads to the possibility of statements such as "The Manchester machine contains 64 magnetic tracks each with a capacity of 2560, eight electronic tubes with a capacity of 1280. Provided it could be carried out sufficiently quickly the digital computer could mimic the behavior of any discrete-state machine. This special property of digital computers, that they can mimic any discrete-state machine, is described by saying that they are universal machines. The existence of machines with this property has the important consequence that, considerations of speed apart, it is unnecessary to design various new machines to do various computing processes. They can all be done with one digital computer, suitably programmed for each case. Contrary Views on the Main Question We may now consider the ground to have been cleared and we are ready to proceed to the debate on our question, "Can machines think? We cannot altogether abandon the original form of the problem, for opinions will differ as to the appropriateness of the substitution and we must at least listen to what has to be said in this connexion. Nevertheless I believe that at the end of the century the use of words and general educated opinion will have altered so much that one will be able to speak of machines thinking without expecting to be contradicted. The popular view that scientists proceed inexorably from well-established fact to well-established fact, never being influenced by any improved conjecture, is quite mistaken. Provided it is made clear which are proved facts and which are conjectures, no harm can result. God has given an immortal soul to every man and woman, but not to any other animal or to machines. I am unable to accept any part of this, but will attempt to reply in theological terms. The arbitrary character of the orthodox view becomes clearer if we consider how it might appear to a member of some other religious community. It is admitted that there are certain things that He cannot do such as making one equal to two, but should we not believe that He has freedom to confer a soul on an elephant if He sees fit? We might expect that He would only exercise this power in conjunction with a mutation which provided the elephant with an appropriately improved brain to minister to the needs of this sort[. In attempting to construct such machines we should not be irreverently usurping His power of creating souls, any more than we are in the procreation of children: rather we are, in either case, instruments of His will providing. I am not very impressed with theological arguments whatever they may be used to support. The popularity of the theological argument is clearly connected with this feeling. It is likely to be quite strong in intellectual people, since they value the power of thinking more highly than others, and are more inclined to base their belief in the superiority of Man on this power. I do not think that this argument is sufficiently substantial to require refutation. There are other, in some respects similar, results due to Church (1936), Kleene (1935), Rosser, and Turing (1937). There may, of course, be many such questions, and questions which cannot be answered by one machine may be satisfactorily answered by another. We are of course supposing for the present that the questions are of the kind to which an answer "Yes" or "No" is appropriate, rather than questions such as "What do you think of Picasso? The short answer to this argument is that although it is established that there are limitations to the Powers If any particular machine, it has only been stated, without any sort of proof, that no such limitations apply to the human intellect. It is no doubt quite genuine, but I do not think too much importance should be attached to it. In short, then, there might be men cleverer than any given machine, but then again there might be other machines cleverer again, and so on. Those who hold to the mathematical argument would, I think, mostly he willing to accept the imitation game as a basis for discussion, Those who believe in the two previous objections would probably not be interested in any criteria. No mechanism could feel (and not merely artificially signal, an easy contrivance) pleasure at its successes, grief when its valves fuse, be warmed by flattery, be made miserable by its mistakes, be charmed by sex, be angry or depressed when it cannot get what it wants. According to the most extreme form of this view the only way by which one could be sure that machine thinks is to be the machine and to feel oneself thinking. Likewise according to this view the only way to know that a man thinks is to be that particular man. A is liable to believe "A thinks but B does not" whilst B believes "B thinks but A does not. The game (with the player B omitted) is frequently used in practice under the name of viva voce to discover whether some one really understands something or has "learnt it parrot fashion. And so on, What would Professor Jefferson say if the sonnet-writing machine was able to answer like this in the viva voce? I do not know whether he would regard the machine as "merely artificially signalling" these answers, but if the answers were as satisfactory and sustained as in the above passage I do not think he would describe it as "an easy contrivance. I do not wish to give the impression that I think there is no mystery about consciousness. There is, for instance, something of a paradox connected with any attempt to localise it. They are ugly, each is designed for a very limited purpose, when required for a minutely different purpose they are useless, the variety of behaviour of any one of them is very small, etc. Naturally he concludes that these are necessary properties of machines in general.
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Some journals have a specific format for additions or corrections; check the author guidelines arteria occipital cheap betapace 40 mg line. In books arrhythmia effects order betapace in india, errata sheets will be printed and included in every book hypertension quizlet order betapace in united states online, and the book itself will be corrected before reprinting. Note that corrections to manuscripts are made in place (not in the margins) and usually need no additional explanation. Authors may be called on to use this proofreading method if the publisher supplies hard-copy (paper) proofs. For example, a carat mark (^) in the typeset text indicates where new words are to be inserted; the words themselves are written in the margin. Avoid using arrows and lines to indicate where corrections go because more than one or two on a page breed confusion. Double underline to make "ord" small capitals Draw a wavy line to indicate bold face. Put dots or short dashes under copy that you wish to retain as it originally appeared. Chapter 2 discusses the parts of a scientific paper; Chapter 3 presents an overview of the editorial process. Chapters in Part 2 address more specialized rules for usage; see, especially, Chapter 9 on grammar, punctuation, and spelling and Chapter 10 on editorial style. Short, simple declarative sentences-that is, sentences that make statements, rather than pose questions, issue commands, or exclaim-are the easiest to write and the easiest to read. They also can place too heavy a burden on the reader to connect the ideas from one sentence to the next. Identity specifications and tests are not included in the monographs for reagent chemicals. Tense Using the appropriate verb tense helps to orient the reader as to the nature of the information. Hyperbranched compounds are macromolecular compounds that contain a branching point in each structural repeat unit. Sentence Modifiers Modifiers made up of phrases or dependent clauses can be added to simple sentences to indicate, for example, cause and effect, or time sequence, or comparison. Several systems that take advantage of this catalysis can be used to create new palladium compounds. A nonrestrictive phrase or clause is one that adds meaning to the sentence but is not essential; in other words, the meaning of the basic sentence would be the same without it. Doctoral students, who often have completed their coursework, apply for this teaching fellowship. It should not be confused with an absolute construction, which modifies an entire sentence. When we added 2 mL of indicator solution, the end point for the titration was reached. Sentence Construction and Word Order Use an affirmative sentence rather than a double negative. Equal grammatical rank means that words are connected only to words, phrases only to phrases, subordinate clauses only to other subordinate clauses, and sentences only to other sentences. Establish parallel construction by using coordinating conjunctions, correlative conjunctions, and correlative constructions. Chapter 4: Writing Style and Word Usage 47 A coordinating conjunction is a single word, such as "and", "but", "or", "nor", "yet", "for", and sometimes "so". Use parallel constructions in series and lists, including section headings and subheadings in text and tables and listings in figure captions. Use "compare" followed by the preposition "with" when differences are being noted. Chapter 4: Writing Style and Word Usage 49 incorrect the alkyne stretching bands for the complexes are all lower than the uncoordinated alkyne ligands. Idioms often used in comparisons are "different from", "similar to", "identical to", and "identical with". The greater acidity of nitric acid relative to nitrous acid is due to the initial-state charge distribution in the molecules. The lowering of the vibronic coupling constants for Ni as compared with Cu is due to configuration interaction. Chapter 4: Writing Style and Word Usage 51 Use the more accurate terms "greater than" or "more than" rather than the imprecise "over" or "in excess of ". On the basis of the molecular orbital calculations, we propose a mechanism that can account for all the major features of alkali and alkaline earth catalyzed gasification reactions. Chapter 4: Writing Style and Word Usage 53 correct I am not sure whether to repeat the experiment. See pp 257 and 264 for the use of "a" and "an" with chemical elements and isotopes. Chapter 4: Writing Style and Word Usage 55 instead of in spite of the fact that in the case of. Such phrases would be appropriate only if you were asked to provide an exact transcript of a speech. Recent style guides and writing guides urge copy editors and writers to choose terms that do not reinforce outdated sex roles. Instead of "man", use "people", "humans", "human beings", or "human species", depending on your meaning. Instead of "man-made", use "synthetic", "artificial", "built", "constructed", "manufactured", or even "factory-made". Blendermann lectronic submission of manuscripts to journals is undergoing significant change as publishers respond to mounting pressure to publish faster, better, and more efficiently. And although scientific research has always been an international activity, journal publishing is increasingly global, with authors, reviewers, and editors contributing from numerous countries and all participants benefiting from electronic communications. If author source files are not of adequate quality for production, then the publication process will be delayed. However, authors can rely on this chapter to guide them through the general process of online submission. Proprietary and commercial editorial systems are frequently designed to accommodate both electronic and paper publication processes. Each package provides authors, reviewers, and editors with submission acknowledgments, decision letters, and review documents transmitted using e-mail; no paper correspondence is needed. They indicate the types of components that are required for online submissions, such as the cover letter, abstract, manuscript document, supporting information, figures, and proposed reviewers. Publishers require these items to be submitted in common word-processing or graphics formats, and author guidelines provide the technical details for preparing these manuscript components.
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The successful adaptation of different cell lines to hypertension with cardiac involvement buy 40mg betapace with amex non-ammoniagenic medium has been described by Hassell and Butler (1990) blood pressure chart by age canada generic betapace 40 mg without a prescription. A mole to heart attack by demi lovato buy 40 mg betapace mastercard mole substitution of glutamine by glutamate was successful for a McCoy cell line and led to normal growth rates after approximately 10 days. Cell yield was increased by 17%, ammonia accumulation was reduced by 70%, and glucose consumption and lactate production both decreased by more than 70%. The authors proposed that the glutamate uptake, and not the glutamine synthetase activity, is responsible for the ability of a given cell line to grow in a glutamine-free medium (McDermott and Butler, 1993). Cells transformed with the glutamine synthetase gene can grow in media supplemented with glutamate instead of glutamine, with a direct elimination/ reduction of ammonia generation, either by glutamine decomposition or metabolism (Paredes et al. Ammonia concentrations in the medium of batch cultures of these cells were below detection levels. This cell line could not be adapted to glutamine-free growth even in the presence of elevated levels of glutamate, so metabolic engineering was the unique alternative. Another strategy for the reduction of ammonia accumulation can be the use of glutamine-containing dipeptides, which hydrolyze slowly in the culture (Butler and Christie, 2004). The supplementation of a glutaminefree medium with dipeptides containing glutamine allows a reduction in the rate of ammonium generation, but this requires the presence of dipeptidases, which may be produced by the cells and may be released into the medium. They Cell metabolism and its control in culture 87 obtained high cell yields in the presence of 6 mM Ala-Gln or 20 mM Gly-Gln, with the final cell yield in Gly-Gln 14% higher than in Gln. The higher concentration of Gly-Gln was necessary for cell growth because of the presence of a peptidase (in the cytosolic fraction of the cells) with a lower affinity for Gly-Gln. Substrate utilization and metabolism was affected by the presence of the dipeptides, particularly with Gly-Gln. The specific consumption rates of glucose and six amino acids were reduced and the accumulation of ammonia and lactate was significantly lower. Amino acid profiles from the cell growth phase for pyruvate medium showed a reduced uptake of serine, cystine, and methionine, an increased uptake of leucine and isoleucine, and a higher release of glycine compared with glutamine medium (Genzel et al. The fraction of metabolic energy obtained from amino acids varies greatly with the type of cell and with metabolic conditions. The transport of amino acids into mammalian cells can be regulated by nutritional, hormonal, or other environmental factors or by changes within cells like transformation. The intracellular or extracellular concentration of amino acids probably has the most profound influence on the efficiency or capacity of transport into animal cells. The importance of amino acids in synthetic media for in vitro growth of mammalian cells has long been recognized, as both a nitrogen donor and a carbon source. Studies on the rates of amino acid uptake have shown generally that glutamine is the most rapidly consumed, followed by lysine, leucine, and isoleucine (Roberts et al. The nutritional requirement for a certain metabolite, however, may also be influenced by the cell population density. For example, serine, cystine, glutamine, and asparagine have been shown to be required at low, but not at high, cell densities (Eagle and Piez, 1962). This occurs in situations when the metabolite is utilized in amounts that exceed the biosynthetic capacity of the cell. At high cell densities, however, the cell culture medium may require supplementation with extra amino acids, to prevent their depletion (Doyle and Griffiths, 1998). Higher eukaryotic cells have lost the ability to synthesize a number of amino acids. These amino acids are generally called essential amino acids, while those that can be synthesized are called non-essential. First, some of the nonessential amino acids are in fact very essential in that they are required for synthesis of nucleotides (glycine, aspartate, and glutamine). The reason why the ability to synthesize these amino acids has been retained may well be that they are indispensable. Secondly, the capability to synthesize them Cell metabolism and its control in culture 89 may be conditional depending on the nutritional situation, the proliferative status, and cell line-specific properties (Doverskog et al. The early work of Eagle (Eagle, 1955) demonstrated a need for 12 amino acids to support the proliferation of strain L mouse fibroblasts in medium containing 0. Glutamine was later added to this list (Eagle, 1959), which also includes arginine, cystine, histidine, isoleucine, leucine, lysine, phenylalanine, methionine, threonine, tryptophan, tyrosine, and valine. The rates at which these and other amino acids are utilized or produced can vary dramatically between cell lines. The relative concentration of amino acids and serum in the culture medium and other conditions of the culture environment will also influence the rates of utilization or production of specific amino acids. Amino acids, whose carbon skeleton can be synthesized de novo in mammalian cells, include serine, glycine, alanine, aspartate, asparagine, and in principle glutamate and glutamine. Although most cells possess glutamate dehydrogenase, it is doubtful if there is any significant net synthesis of glutamate through this enzyme in cultured cells. Glutamine, proline and ornithine are all synthesized from glutamate (Doverskog et al. The amino acids that can be synthesized by a cell depend upon the strain-specific profile of biosynthetic enzymes. Another example of a strain-specific difference is the ability to synthesize glycine. This enzyme, which converts serine to glycine and tetrahydrofolate-bound single-carbon units, is present both in the cytoplasm and mitochondria. The mitochondrial isoenzyme activity may be absent in these partial auxotrophs, which are self-supporting in single-carbon units through the cytoplasm enzyme activity, but they need glycine from the medium for protein synthesis. In contrast to mammalian cells, insect cells are much more flexible in their amino acid metabolism (Ferrance et al. The capability of synthesis of a certain amino acid may be conditional, and depends on the availability of carbon precursors and nitrogen donors. For example, a glutamine-free medium for mammalian cells may have to 90 Animal Cell Technology be supplemented with aspartate and/or asparagine (Bebbington et al. Another example of conditional biosynthesis is the formation of cystine (from methionine, the cystathionine pathway) in Sf9 cells, which appears to be regulated in relation to the proliferative status of the cells (Doverskog et al. It has been recognized that certain cells have a specific requirement for an amino acid, for example, serine for lymphoblastoid cells (Birch and Hopkins, 1977). This may be due either to the inability of the cells to make an amino acid, or because the amino acid is decomposed in the medium. The concentration of amino acids usually limits the maximum cell concentration attainable, influences cell survival and growth rate, and can affect the synthesis of certain proteins. A too low concentration of an amino acid can result in rapid depletion from the medium, and is thus ``limiting,' whereas a too high concentration can be inhibitory. The sulfur-containing amino acids, methionine and cystine, are also rapidly consumed by cells in culture (Lambert and Pirt, 1975; Butler and Thilly, 1982). It is suggested that a function of glutamine uptake and glutamate formation is to allow cystine uptake by glutamate exchange into the culture medium (Bannai and Ishii, 1988). Certain amino acids often accumulate in the culture medium during batch growth with surplus glucose and glutamine (Griffiths, 1971; Lanks and Li, 1988; Duval et al.
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The skin will also undergo changes and begin to hypertension genetics order betapace 40mg amex take on a reserve of fatty tissue arrhythmia quizlet 40 mg betapace visa. In autumn the animal will grow a new winter coat and the skin will again change and the fatty tissues in the skin dissolve pulse pressure 82 generic betapace 40mg fast delivery. These seasonal changes affect the quality, colour and grade of pelt which in turn affect the value. It was not until the latter part of the nineteenth century that commercial raising and breeding of furbearing animals was introduced. Many species breed only once a year and only certain species do well in captivity in pens. Animals that are confined are also more prone to disease, parasites and bacterial infections. They also needed an understanding of genetics and a planned and well-recorded breeding programme in order to have the likelihood of breeding good stock and not to in-breed too closely and thus have unplanned mutations and weaknesses occur in successive generations. Today with all the benefits of modern science, zoos are still struggling with breeding certain animals in captivity and are not always successful. For species that are still hunted, population and habitat management ensures this viability. This is achieved by scientific monitoring by professional wildlife biologists and governmental regulations which allow for a certain number of a species to be harvested each year without causing threat to the survival of that species. Other species farmed on a smaller scale include nutria (coypu, Myocaster coypus), chinchilla (Chinchilla lanigera), fitch (polecat also known as foul marten, Mustela putorius), sable (Mustela zibellina), and finnraccoon (Tanuki Nyctereutes procyonides). From a scientific point of view, fur animals that have been domesticated (farmed) for more than ten generations are considered to be so far genetically removed from their ancestors that they must be treated as a fully domesticated subspecies. Fur farming is regulated and in the European Union, Council Directive 98/58 sets down rules covering the welfare of farmed animals, including furfarmed animals. Directive 93/119 deals with the slaughter and killing of fur and other farmed animals. In North America, fur farmers also follow strict codes of practice and conform to provincial, state or national animal welfare and other regulations. In November 2000 a bill to ban fur farming in England was passed by Parliament and came into 146 Conservation of leather and related materials force at the end of 2002 forcing the 13 mink farms to close down. It is useful to the conservator to know what are the most likely species which may be found in fashionable dress and when they were used. It does not, however, preclude any of the 100 or so species appearing in an object but it may be relevant to know when a particular species was fashionable and available in order to help pinpoint the likely date of the object concerned, and how the fur may have been processed. To a fur-skin processor (or fur-dresser) and to the furrier concerned with the manufacture of fur garments, etc. Included in these are sheep and lambs which by convention are commonly described as possessing wool. During fur processing sometimes these guard hairs are removed to expose the soft underfur. There are few differences across the continents in the manner in which raw skins were and are commercially prepared. It reads: the skin of the kid thou shalt feed with the milk of a yellow goat, and with flour; thou shalt anoint [it] with pure oil, ordinary oil, and the fat of a pure cow. Thou shalt dilute alum in pressed grape juice, then fill the surface of the skin with gall nuts of the tree-cultivators of the Hittites. It is logical to assume that as the Babylonians used and traded furs, a recipe similar to this would have been used for their preparation and dressing. By Roman times a distinction was made between tanners (coriarii) and fur-skin dressers (pellioni) and it was recognized that each was a separate distinct skill and process. In addition to dressing, pellioni included in their activities the making up of certain items and a general trading in fur-skins. Tanners concentrated on bark tanning and tawyers concentrated on those skins for which the use of alum and oil was more suitable. By medieval times the furrier combined the activities of fur-skin dresser with that of manufacture of garments and trimmings. The guilds covered many branches of a particular trade and it was considered fraudulent practice to dye fur-skins. However this practice did not include trimmings matched to a garment and dyed to red, blue or green, only the practice of dyeing to cover up natural faults in a skin, light spots or patches with the intent to deceive. Black dyeing was especially forbidden and remained so until the seventeenth century. According to the records of the Skinners Company alum and oil were used for fur dressing but by 1593 alum dressing had been partially replaced by oil. The method used for preparing fur-skin was to stretch the skin on a frame and sponge the flesh side several times with a solution of alum and salt, or oil. Furs and furriery: history, techniques and conservation 149 Lightweight fur-skins such as squirrel, beaver and fox were first greased with oil, butter or other fatty substances and trampled with bare feet in a barrel until the skin was rendered pliable. In the seventeenth, eighteenth and nineteenth centuries dyeing was less restricted although still controlled. Black and brown were dyed onto furskins using plant products such as logwood, sumac and gallnuts together with mineral substances such as verdigris, iron filings, alum, or copper scale. By the mid-nineteenth century demand for furs had increased to the extent that large factories employing hundreds of workers were set up in Europe to process furs rather than small workshops with only a small number of craftsmen. Methods of dressing were still dependent on alum, salt and natural fats, and for dyeing upon vegetable products and metal salts. The fur seal which became popular around 1847 was dyed by repeated brush applications of dye in order to preserve the leather from damage by the strong dye, the composition of which was based on that used in the French silk industry (Kaplan, 1971). By the end of the nineteenth century mechanization of some processes had taken place. The discovery of synthetic coal tar dyes by Perkin in 1856 had no immediate application in fur dyeing and it was not until the end of the century when oxidation dyes were patented that coal tar derivatives could be applied to furs. The use of para-phenylene diamine enabled a greater range of colours to be obtained on fur. The mammalian skin is constructed mainly from a series of interlaced protein fibres of which the four main types are described as follows: 1. Collagen fibres forming the bulk of the grain and corium, and constitute what is generally known as the pelt or leather. The fibre, described variously as hair, wool or fur depending on the animal type, is composed of the protein keratin. The fur-skin processor deals with the keratin fibres and the three other types of fibrous proteins in an inseparable form, his aim being to maintain and enhance not only the properties of the fur fibre but equally the skin fibres. The function of these keratin fibres is to serve as a protection for the skin and to insulate against weather and other hazards of the environment. This is true of warm climates as much as cold; fur-covered tropical animals are insulated against strong sun and heat. The air trapped between the fibres helps to form an efficient insulating layer, preventing rapid loss of heat which is why animals from cold regions are more fully furred than those from warm climates. Certain species of mammals, such as pigs, have very little hair and in this case a layer of fat under the skin plays the role of temperature regulator.
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The lath and surrounding leather were covered in cling film to blood pressure 9060 best betapace 40mg keep them dry while the new leather was thoroughly wetted and moulded around the lath pulse pressure greater than 70 discount 40mg betapace free shipping. When dry arteria hyaloidea persistens discount betapace express, the new leather held its shape and was trimmed to size and bonded into place with the 272 Conservation of leather and related materials Figure 23. The original had a very heavy layer of paint on it and when the new leather was painted to match the grain texture showed through. The thick paint on the original was therefore replicated by working solid Beva into the surface of the new leather with a heated spatula. The weak sections of the handle were reinforced by inserting new vegetable-tanned calf between the layers and then replacing the missing stitching using the original holes. The bucket was to go into an environment where it was likely to be handled by untrained staff so it was very likely that it would be lifted by its handle. The original disintegrating loops used to hold the handle to the main body were so damaged that it was felt a satisfactory, strong repair was impossible. As a result they were replaced using new leather stitched into place through the original holes. Although the work was not for a museum, the owner still asked for a minimum level of intervention consistent with the handling demands of the object. The exterior of the bucket was lightly finished with Renaissance micro-crystalline wax polish and the handle was lightly dressed with Pliantine (Figure 23. It is made from degrained, vegetable-tanned cattle hide, except for the hatband which is vegetabletanned sheepskin. The main body of the helmet was in good condition, but the brim was in poor condition (Figure 23. These soft sponges crumble with the dirt sticking to the fragments which are brushed and vacuumed away. However, they need to be used with care, as on a fragile surface they can cause damage and remove flakes. The red rot on the rim and hatband was treated with a 1% solution of aluminium alkoxide in mineral spirits. This latter process is important, as if there is an excess left on, an insoluble white deposit can remain on the surface. The fragile rim was consolidated with a 3% solution of Pliantex, a flexible acrylic resin, in toluene. Although this consolidated the body of the leather, it did not hold the flakes in place. Because it is in the form of a dispersion rather than a solution, the adhesive does not soak into the leather. The hole in the rim was filled with new vegetabletanned calfskin coloured with Sellaset dyes. As this was not a strong join it was reinforced with suitably coloured Stabiltex (polyester) fabric. This was first applied to the Stabiltex, which was then cut to size and applied to the join. Stabiltex was chosen for its strength and relative transparency, which made a discreet repair possible. The areas on the rim where the grain surface had been lost were coloured with Sellaset dyes. This was applied locally with a brush to tone the damage in with the surrounding colour. A final light finish was applied to the entire helmet using Renaissance microcrystalline wax polish (Figure 23. The vegetable-tanned leather had then been moulded over the core, presumably while wet. It had been handled over the years, leaving a smooth surface and some slight soiling. The main damage was to the end of the tail where a section of the leather and core were missing (Figure 23. The surface of the original plaster was friable so it was consolidated with 5% Paraloid B72 in toluene to give a sound surface for the Milliput to bond to. The Milliput was chosen as it bonded well to the original core and it is easy to model as it cures. The areas where the leather was missing were re-covered with new vegetable-tanned calfskin. It was even possible to work it into the claws of the lion so that they showed on the front. The whole lion was lightly cleaned and polished with Renaissance micro-crystalline wax polish. Pigments in wax were chosen, rather than dye, as it was a surface patination that was required rather than an in-depth colouring of the leather (Figures 23. The leather was attached with dome-headed brass tacks arranged to give a simple decorative border. The windows, which could be lowered on braided straps, were framed with a smooth finished gilt timber subframe. Around the windows and along the top, just beneath the roof, there was a carved gilt wood trim. The patent leather finish was in good condition except for a few vertical lines on the sides, apparently caused by a liquid running down. However, it had split in a number of places both on the flat sides and on the concave segments of roof. The gilt areas were in good condition except for some fairly light soiling and some worn areas on the highest points. The textiles on the inside were heavily soiled and one of the straps for lowering the window had broken at the point where it joined the sash frame. The method chosen for each repair depended on two factors: the availability of access to the back of the leather, and whether or not the leather was flat, as on the vertical surfaces, or curved, as on the roof. The only areas where there was any access to the back of the leather without going through the split were in those places where the interior fabric had come away partially. This did not give full access, but it did allow some unwanted materials used during the repair work to be removed when the work was finished. The sandwich was made up with stiff card on the outsides, two layers of silicone release paper and the repair material in the middle. A tracing of the split was made on Melinex and this was used to pierce pairs of holes along the line of the split. The back piece of card, silicone release paper and the repair material were inserted through the split and then drawn up into position using the threads. Adhesive was then inserted between the repair material and the back of the leather. Once the position was correct, the outer silicone paper and card were slid into place down the thread which was cut and then retied close to the leather. Wedges were then inserted in the loops to pull them tight and hold the sandwich tightly together (Figure 23. In places where the card on the inside would drop into an inaccessible space, it was found helpful to tie a thread to the corner prior to insertion and bring this out through a suitable gap.
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There are many publications available in conservation literature on the topic of pest management including Florian (1987) xeloda arrhythmia cheap 40mg betapace, Child and Pinniger (1993) heart attack from weed purchase 40 mg betapace free shipping, Hillyer and Blyth (1992) prehypertension 34 weeks pregnant cheap 40mg betapace visa, Pinniger (1994, 1995), and Norton (1996) the opinion expressed by writers on furs and furriery is that moths are the most damaging risk to furs and that cold storage balanced with a controlled atmosphere is the best remedy (Sachs, 1922; Rosenberg, c. If the atmosphere is maintained at a high humidity in very low temperatures, moisture will condense into a frost on the garment; but if a dry atmosphere is maintained the cold air will absorb the little remaining moisture causing the skins to dry out and become brittle. It is therefore essential to balance the environmental conditions when furs are kept in reduced temperature storage and specialists in this field should be consulted if this type of storage is to be provided. Maintenance of furs in reduced temperature storage of no more than 8°C (50°F) was recommended by Bachrach as far back as 1930 but few museums have the facility for this. Clothes moths (Tineola biselliella and Tinea pellionella) will not breed or feed below about 10°C. However, this temperature may not kill larvae for a long time as they probably starve to death. The larvae of carpet beetles (woolly bears) may be more tolerant but the adults will not breed at 8°C. Until the 1980s treatment for pest infestations was carried out using methyl bromide and ethylene dioxide but since then deep freezing of infested material has become one of the accepted methods and is a straightforward procedure to carry out. Deep freezing to temperatures of 30°C for a period of 72 hours and then repeating the procedure is currently the accepted method of dealing with insect infestations in museums (Florian, 1987). In 2003 it became accepted that it is unnecessary to carry out the second freezing procedure. Anoxic treatments using nitrogensaturated atmospheres have also been regularly and successfully in use for a number of years (Newton et al. An alternative for whole collections and large quantities of fur items is to treat them using the thermo-lignum process3 (Thomson, 1995). Beating the fur from time to time with sticks is advocated by furriers and is a treatment that goes back to medieval times. William Jurden, Skyner to Queen Elizabeth I, had the essential task of regularly airing and beating the furs and fur-trimmed garments to keep them free of dust, moths and fleas (Arnold, 1988). Although beating is still carried out by furriers today as part of maintenance it is not a suitable treatment for fragile aged skins or museum objects. An exception was an exhibition held at the Museum of London, November 2000 entitled Stolen Skins? The first involves raising the temperature gradually from an ambient temperature of 20°C up to 50°C over a period of 11 hours, holding this temperature stable for two hours then allowing cooling back to ambient temperature of 20°C over 10 hours. During this time the relative humidity is strictly controlled and raised from 50 to 60% and back again. The second process involves flooding the chamber with argon to give an atmosphere of less than 1% oxygen, then heating to 38°C over a period of 8 hours, holding at that temperature for 3 days and then cooling over a further 8 hours. During this treatment 168 Conservation of leather and related materials London, 11 December 2000, pp. Both methods are useful for fur-skins but it is the second method that could be useful for degraded fur-skins with very low shrinkage temperatures. Delany, Mrs (1861) the Autobiography and Correspondence of Mary Granville, Mrs Delany. Seminar organized by Conservators of Ethnographic Artefacts held at Museum of Furs and furriery: history, techniques and conservation Indian, Smithsonian Institution. Seminar organized by Conservators of Ethnographic Artefacts held at Museum of London, 11 December 2000, pp. In Leather and Fur, Aspects of Early Medieval Trade and Technology (Esther Cameron, ed. A fair proportion of these will be made from exotic leathers, distinguishable from mammalian leather by their interesting surface texture, such as scales or quill pips and by their fascinating natural markings. These skins were mostly dried and made supple by scraping the flesh side and applying various animal fats, methods that preserve the skin without imparting a tannage. The supply of exotic skins in bulk, especially to tanners in the United Kingdom, started towards the end of the nineteenth century from countries within the Empire situated within tropical regions. Rain forests, savannas, lakes, rivers and the sea contained wild species with skins large enough to be commercially usable in the manufacture of leather goods. The import of East Indian tanned buffalo hides and goatskins provided the framework for the trade in exotic skins to commence, exported either air-dried, in a raw, wet-salted condition or pre-tanned in a local tannery. France initially obtained its supplies of exotic skins from its colonial empire within Africa where the rivers and lakes yielded an abundant supply of crocodile skins. The expansion of the trade soon necessitated searching other areas apart from Africa and the Far East for supplies. Exotic skins were imported from South and North America, and from other tropical or subtropical areas. For any skin to be commercially usable it has to satisfy the following requirements: (a) the available area of skin must permit a manufacturer of finished leather articles to cut the components with a minimum of waste. It follows that the larger the leather article, such as a handbag, the larger the exotic skin has to be, except where a design is used permitting smaller pieces to be stitched together. Reptile skins with their tightly woven fibre structure are immensely strong, particularly with a skin which is relatively thick, such as crocodile. The skins of 170 the tanning, dressing and conservation of exotic, aquatic and feathered skins 171 birds, such as ostrich or emu, have a looser fibre structure as have fish which results in a lower tensile strength. Exotic skins have good resistance to scuffing with lizard skins among the best compared to leather made from domestic animals. As such it must be available in fashionable colours and in the desired surface finish, such as bright, shiny, metallic, matt, or dull, such as suede. The tanner has limitations imposed on him by the nature of the skin as well as fashion demand, with crocodile leather for handbags, invariably finished by glazing, giving a brilliant finish. Bird and fish skins were nearly always finished matt after tanning, their fibre structure being too loose to be glazed, but modern finishing systems allow more shiny effects if not the brilliant results obtained from glazing. These are widely used for American cowboy boots with the wide belly scales running down the centre of the boot and vamp. Rattlesnakes (genus Crotalidae), in the raw, dried state, are also used, mostly belly-cut in the manufacture of these boots. Crocodiles and alligators are very heavily ossified on their backs which makes this part of the skin impossible to tan and soften. For this reason only the sides and belly are used in the tanning process (Figure 16. Lighter ossification is also present in the belly of some species which, if not too severe, can be softened by the tanner with acids. After thoroughly soaking air-dried skins, they are immersed in a solution of sodium sulphide to remove the keratinous surface layer of the scales, which is the equivalent to the hair and epidermis of mammalian skin. After bating and pickling with acid follows the important bleaching process which removes, Figure 16. With the demand mostly for glazed reptile leathers, the traditional chrome tannage cannot be used or only as a pre-tannage on crocodile leathers.
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The low percentage reflects consistent and correct use; the high blood pressure medication problems purchase betapace 40 mg with amex, less consistent and correct use arrhythmia headaches betapace 40 mg otc. Many of the reasons that women do not arteria circumflexa femoris lateralis order betapace pills in toronto, or are unable to, protect themselves against unplanned pregnancy are not so much personal as social, cultural or economic. In some traditional agrarian societies, the concept of planning for or preventing pregnancy is not yet widely accepted, although attitudes are steadily changing. In other settings, women who would like to achieve control over their childbearing do not have access to the contraceptive supplies or family planning services they need, because contraceptives are too expensive, supplies are erratic, or services are difficult or impossible to get to. And in other countries, male partners or motherswoman is potentially capable of giving birth about 12 times, in-law with a great deal of power in the extended family diseven if she breastfeeds each baby for one year. So if the avercourage or forbid women from obtaining and using birth age woman is to have a small family and control methods. This is particularly avoid any unplanned pregnancies, she true when very young brides have not Chart 2. Others hear pregnant until they reach menopause; unfounded or exaggerated rumors but as long as they remain sexually about such side effects and are deterred active and capable of conceiving, in no from even trying these methods. The proportion of women who these reasons are not frivolous or describe a recent birth as mistimed or unconsidered. Rather, they demonstrate many of the diffiunwanted is striking everywhere, but it is particularly high culties that beset women in all walks of life who are trying in Kenya and Zimbabwe, in the Philippines, in the four to juggle competing roles and competing responsibilities and representative Latin American countries and in Japan 18 trying to adapt to changing societal expectations. More than two in four women live under laws that permit abortion on broad grounds, but even these laws impose certain limitations on the procedure. Since the 1950s, many developed and some developing countries have liberalized their abortion laws, usually to minimize morbidity and mortality from clandestine procedures, but this trend has been slow in Africa and has hardly touched Latin America. Where abortion is severely restricted by law, prosecution of women for obtaining abortion is rare but persists in a few countries. While most well-off women everywhere can obtain a safe abortion-either in their own country or abroad-poor women often lack access to safe services, even when legally entitled. One of the most significant considerations affecting the circumstances she encounters will be whether the law permits or prohibits abortion where she lives. The remaining 61% of women live under more liberal laws: 20% in countries that permit abortion for socioeconomic reasons, as well as for the narrower grounds described above, and 41% in countries where women may obtain the procedure without being required to give a specific reason (Chart 3. Nevertheless, the number of people in the developing world who are living under liberal laws (2. Even in Countries with Liberal Laws, There Are Usually Some Restrictions In the 55 countries where abortion is permitted on general grounds or for socioeconomic reasons, the law usually stipulates some conditions. Of the countries with liberal laws, all but five (Canada, China, North Korea, Vietnam and Zambia) specify some limitation on how late in the pregnancy a termination may be performed. The limit is 12 weeks in 36 countries, 1422 weeks in eight and 24 weeks or fetal viability (generally considered to be 2324 weeks of pregnancy) in six. Most countries whose laws include gestational limits usually permit abortion later in pregnancy in some circumstances, or with additional requirements (such as approval by two physicians, rather than one). Countries that permit abortion on socioeconomic grounds usually interpret these laws quite liberally. In Great of medical facility in which abortions must be performed Britain, Taiwan and Zambia, for instance, the law takes and the type of health professional permitted to perform into consideration the effect that a continued pregnancy them. In Great Britain, India and South Africa, for example, might have on the children a woman already has. Most countries require aborhealth generally must show that their tions to be performed by qualified life would be threatened or that they physicians, but where physicians are would sustain serious or permanent not available, midwives or other regisChart 3. Cuba, Denmark, % 41 life of the pregnant woman or of France, Italy, Norway, Turkey and avoiding grave and permanent harm to most countries in Eastern Europe, for her health. And in women to obtain abortions on mental Turkey, an adult woman must have health grounds are wide-ranging. Depending on the country, these might However, in France, if a minor include psychological distress as a result wishes to have an abortion without 1. Some countries have laws aimed at ensuring that women fully understand the proceCountries with Restrictive Laws May Allow Exceptions dure and its ramifications, and sometimes at discouraging her from following through on her decision to obtain an Some countries permitting abortion only to safeguard a abortion. For example, pregnancy terthe procedure and possible alternatives to abortion; she must mination may be permitted in these countries if the woman then wait six days before having the abortion. In Germany, a has been a victim of rape or incest, or if it can be shown woman is required to have counseling that has the stated that her child will be born with serious defects. Brazil, Mexico, Panama and Sudan, which have highly restrictive aim of dissuading her from ending her pregnancy. And the Democratic Some Requirements Can Be Broadly Republic of Congo and Panama permit the procedure when Interpreted the fetus is impaired (Table 3a, page 23). For abortions are performed or reported under the exception example, a woman seeking an abortion on socioeconomic for fetal impairment. Women May Need Help Navigating Complex Legal Systems completed, the pregnancies of many eligible women exceed the gestational limit for rape victims (12 weeks). Seven other hospitals in Brazil now likely than those in developed offer similar services to rape victims. Cautious medical attitudes toward Restrictive abortion laws are difficult to abortion, combined with often dauntenforce because in the absence of a ing procedural requirements, are likely complaint, police and judicial authorito make the approval process complities do not easily learn of violations. Even if Even if they do, they often have diffiwomen know that the law makes an culty locating witnesses willing to testi1. Furthermore, sometimes the ambiFor women of any age or social guity in the language of an abortion background, the psychological obstacles are also likely to be formidable. For example, not all laws prohibiting aborwoman seeking a legal abortion after tion after a particular point in gestashe has been raped is forced to relive tion specify how to determine the the horror of her experience with family members or friends, possibly with a length of gestation. The fluidity or vagueness of some mental health All of the above plus the city of Sгo Paulo was the first in legislation helps explain why, even in socioeconomic grounds the country to establish an official hoscountries with the most restrictive Without restriction as to reason pital service to provide this option. In a small number of countries, however, large numbers of During its first three years of operation, the program people have been imprisoned for having or inducing aborserved about 200 women. Some countries that usually permit abortion only on narrow grounds also allow it in instances of rape, incest or fetal impairment. Between 1985 and 1997, 10 developed and nine developing countries with populations of more than one million reduced restrictions on abortion. Of these, 12-including Cambodia, Mongolia and South Africa in the developing world-made first-trimester abortion available without restriction as to reason. Following the pattern of earlier years, only a small proportion of these ended in acquittal, one in seven resulted in a conviction and the great majority were suspended with no decision reached (Table 3b). While the exact number of Nepalese women who have served prison % by outcome Year 1993 1992 1991 1985 1980 Number of prosecutions 245 219 218 396 550 Conviction 15 18 11 17 12 Acquittal 3 5 4 2 1 No decision Total 82 77 85 81 87 100 100 100 100 100 Note: In some instances, women are tried in groups of up to six; therefore, a single prosecution may involve several women. S H A R I N G R E S P O N S I B I L I T Y: W O M E N, S O C I E T Y A N D A B O R T I O N W O R L D W I D E 23 very restrictive abortion laws (Mexico12 and Brazil,13 for example) to encourage political and public debate about the possibility of legal reform. In fact, advocates of abortion law reform in Latin America urge removing the matter from the jurisdiction of national penal codes and making it a public health issue. Year 19971 19951996 19941995 19931994 19921993 Number jailed for abortion 80 76 89 73 82 % of all women jailed 20 18 21 18 20 1. Movements supporting or opposing abortion law reform can come to the fore and then recede again quite rapidly, depending on local events (such as a visit by the Pope, publicity given to a sudden rash of abortion-related deaths or police raids on clandestine abortion clinics) and on the relative importance attached to other pressing social issues.
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The white powder is best stored in an inert atmosphere in the dark in sealed ampoules after drying in vacuo over P2O5 at 34o heart attack demi lovato mp3 order betapace from india. It is readily oxidised in air and is best kept as the more stable trilithium salt [Moffat & Khorana J Am Chem Soc 83 663 1961; see also Beinert et al blood pressure levels generic betapace 40 mg. It crystallises in red needles from pet ether (b 40-60o) and sublimes at high vacuum at a bath temperature of 46-48o [Ashley et al heart attack piano cheap betapace 40mg on line. It is a red oil which can be purified by chromatography on SiO2 plates and eluted with Et2O/hexane. Coenzyme Q9 (U b i q u i n o n e - 9, 2,3-dimethoxy-5-methyl-6-[3,7,11,15,19,23,27,31,35nonamethyl-hexatriaconta-2t,6t,10t,14t,18t,22t,26t,30t,34-nonaenyl]-1,4-benzoquinone) [303-97-9] M 795. The yellow crystals are purified by recrystallisation from pet ether and by chromatography on SiO2 plates and eluted with Et2O/hexane. Helv Chim Acta 42 1278 1959; Morton Biochemical Spectroscopy (Adam Hilger, London, 1975) p 491]. Alternatively, an acetone solution on alkalifree alumina has been used, and eluting with acetone [Nicholls & Tarbell J Am Chem Soc 7 5 1104 1953]. J Biol Chem 253 1121 1978, Nakamura & Ableles Biochemistry 24 1364 1985, Beilstein 18/3 V 145]. Crotaline (monocrotaline, 12,13-dihydroxy-(13 -14 H)-14,19-dihydro-20-norcrotalanan11,15-dione) [315-22-0] M 325. Cytosine crystallises from H2O as the monohydrate 1 2 which loses water on heating above 100o. Purify it by D 589 310 recrystallisation from H2O (hydrated crystals; solubility of the monohydrate is 1. Biochim Biophys Acta 14 456 1964, Todd & Ulbricht J Chem Soc 3275 1960, Lee et al. The crystals are not hygroscopic like the amorphous powder; however, both forms are soluble in H2O but the amorphous solid is about 10 times more soluble than the crystals. When crystallised from hexane, or other solvents, the higher melting form with m 71. When the lower melting forms are kept at their melting temperatures for a while, they are converted to the higher melting form. The pure individual prenol should run as an entity on a chromatogram on paraffin impregnated paper, with acetone as the mobile phase. Phytochemistry 14 1549 1975, Takemoto & Diago Arch Pharm 293 627 1960, Beilstein 22/4 V 371. Purify (-)-ephedrine by vacuum distillation (dehydrates) and forms waxy crystals or granules, and may pick up 0. It gradually decomposes on exposure to light and is best stored in an inert atmosphere in the dark (preferably at -20o). It is a cyclic D Purification of Biochemicals - Miscellaneous Compounds 679 (-)Ephedrine hydrochloride [50-98-6] M 201. The free base separates as white hygroscopic crystals m 52-55o, which are recrystallised from pentane. The free base is unstable in air and light, but the hydrochloride is more stable and best stored as such. It is a muscle relaxant with a curareD like-action and is more active than the -isomer. It recrystallises from H2O to form hydrated crystals which melt at ca 135-140o, resolidifies and melts again at 190-193o. The melting point after drying at 56o/8mm is that of the anhydrous material and is at 137-140o. Larger amounts can be precipitated from water as the uranyl complex by adding a slight excess of uranyl acetate to a solution at pH 6. It can also be purified by adsorption onto an apo-flavodoxin column, followed by elution and freeze drying. If paper chromatography indicates impurities, then recrystallise it from hot H2O or from dilute acid [Walker et al. Fructose-1,6-diphosphate is best purified via the acid strychnine salt pK 3 4 which is stable for several months. A neutral D Purification of Biochemicals - Miscellaneous Compounds 681 solution of the salt keeps well in a frozen state for over several months. Arch Biochem 3 33 1943, Sable Biochemical Preparations 2 52 1952, Stumpf J Biol Chem 182 261 1950]. The barium can be removed by passage through the H+ form of a cation exchange resin, and the free acid is collected by freeze-drying. The 6-phosphate hydrolyses more slowly than the 1-phosphate and considerably slower than pyrophosphoric acid (102 times) and triphosphoric acid (103 times). Alternatively dissolve 90g of it in 700mL of 2O, filter and cool (ca 94% recovery). Purify the fragrant smelling geranyl acetate by fractional 15 D distillation at as high a vacuum as possible. Two litres of a 5% aqueous solution of the phosphate are purified by adjusting the pH to 3. Glycerol phosphocholine can be recovered from the complex by dissolving it in H2O (2% solution) and passing it through an ion-exchange column (4. It is hygroscopic and must be handled in a H2O-free atmosphere [Tattrie & McArthur Biochemical Preparations 6 16 1958, Baer & Kates J Am Chem Soc 70 1394 1948, Acta Cryst 21 79, 87 1966]. It crystallises from Hematoxylin (±-11bc-7,11b-dihydroindeno[2,1-c]-chromen-3,4-6ar-9,10-pentaol) [517-28-2] M 302. Hematoxylin recrystallises from H2O (as trihydrate) in whiteyellow crystals which become red on exposure to light and then melt at 100-120o. R -(-)-2-Hydroxy-3,3-dimethyl- -butyrolactone (3-hydroxy-4,4-dimethyl-4,5-dihydrofuran2-one, D-pantolactone) [599-04-2] M 130. Recrystallise the lactone from D D Et2O/pet ether, diisopropyl ether or *C6H6/pet ether and sublime it at 25o/0. J Am Chem Soc 62 1779 1940, Bental & Tishler J Am Chem Soc 68 1463 1946, Beilstein 18/1 V 22. The dihydrate is efflorescent and becomes anhydrous when heated 684 Purification of Biochemicals - Miscellaneous Compounds at 100o. A further purification step is to convert it to the monocyclohexylammonium salt by passage through a column of Dowex-50 (cyclohexylammonium form) ionexchange resin. Lecithin from hen egg white is purified by solvent extraction and chromatography on alumina. It is suspended in H2O and kept frozen until required [Lee & Hunt J Am Chem Soc 106 7411 1984, Singleton et al. For purification of commercial egg lecithin, see Pangborn [J Biol Chem 188 471 1951]. J Chem Soc 4219 1952, Albert & Wood J Appl Chem (London) 2 591 1952, Pfleiderer Chem Ber 9 0 2631 1957]. It forms yellow needles from cyclohexane or hexane and has been distilled at high vacuum; and sublimes at ~90o and very high vacuum.
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These readily measurable "disease" phenotypes are sensitive to heart attack demi lovato purchase betapace without a prescription temperature prehypertension 139 buy cheap betapace line, gene dose heart attack low blood pressure discount 40mg betapace visa, and sex. The genetic dominance of disease-specific phenotypic variability in our model of misfolded human proinsulin makes this approach amenable to genome-wide association study in a simple F1 screen of natural variation. M an important resource for investigating the genetic underpinnings of continuously varying quantitative traits (Palsson and Gibson 2004; Telonis-Scott et al. Numerous models of human disease have been established in the fly (reviewed in Pandey and Nichols 2011), including transgenic models of diseases ranging from neurodegeneration/retinal degeneration (Bilen and Bonini 2005; Ryoo et al. Success with genetic screens to identify suppressors and enhancers of disease when mutants are overexpressed in a developing tissue, such as the eye-antennal imaginal disc, suggested to us that it might be possible to generate a fly model of misfolded insulin-associated diabetes. Here, we explore an approach for studying epistasis in humans using a Drosophila melanogaster model of neonatal diabetes mellitus. Finally, we evaluated allele-specific expression difference between the two major sfl-intronic haplotypes in heterozygtes. The results showed significant heterogeneity in marker-associated gene expression, thereby leaving the causal mutation(s) and its mechanism unidentified. Drosophila can provide genetic insights relevant to human biology and disease, owing to the conservation of fundamental cellular and developmental processes. We constructed a fly model of protein-misfolding disease, by creating a transgene of a diabetes-causing, human Copyright © 2014 by the Genetics Society of America doi: 10. This misfolded proinsulin protein causes the loss of insulinsecreting pancreatic beta cells and diabetes in humans and mice (Stшy et al. When misexpressed in the Drosophila eye imaginal disc, it disrupts eye development, resulting in a reduced eye area in adult flies (Park et al. The F1 lines displayed a wide, nearly continuous, range of heritable eye-degeneration phenotypes, suggesting a polygenic basis for this genetic background variation (Park et al. It has been suggested that sweet taste receptors in these nongustatory tissues may play a role in systemic energy balance and metabolism. Smaller adipose depots have been reported in T1R3 knockout mice on a high carbohydrate diet, and sweet taste receptors have been reported to regulate adipogenesis in vitro. To assess the potential contribution of sweet taste receptors to adipose tissue biology, we investigated the adipose tissue phenotypes of T1R2 and T1R3 knockout mice. Here we provide data to demonstrate that when fed an obesogenic diet, both T1R2 and T1R3 knockout mice have reduced adiposity and smaller adipocytes. Although a mild glucose intolerance was observed with T1R3 deficiency, other metabolic variables analyzed were similar between genotypes. In addition, food intake, respiratory quotient, oxygen consumption, and physical activity were unchanged in T1R2 knockout mice. Although T1R2 deficiency did not affect adipocyte number in peripheral adipose depots, the number of bone marrow adipocytes is significantly reduced in these knockout animals. Finally, we present data demonstrating that T1R2 and T1R3 knockout mice have increased cortical bone mass and trabecular remodeling. This report identifies novel functions for sweet taste receptors in the regulation of adipose and bone biology, and suggests that in these contexts, T1R2 and T1R3 are either dependent on each other for activity or have common independent effects in vivo. These receptors are reported to function as obligate heterodimers to provide input on the caloric and macronutrient content of ingested food. However, sweet taste receptors have been identified in an increasing number of extra-gustatory tissues , often regulating metabolic processes . In pancreatic b-cells, sweet taste receptors act to augment glucose-induced insulin secretion in response to artificial sweeteners  and fructose . In addition to effects on insulin and incretin secretion [10,13], sweet taste receptors may also have metabolic roles in adipose tissue. Masubuchi et al reported that T1R2 and T1R3 are expressed in 3T3-L1 cells, and that T1R3 is induced during differentiation and mediates inhibition of adipogenesis by artificial sweeteners . Our group also observed that T1R2 and T1R3 are expressed throughout adipogenesis; however, in our hands, saccharin and acesulfame potassium enhance adipogenesis and suppress adipocyte lipolysis through a mechanism independent of both T1R2 and T1R3 . Mice deficient in leptin action are obese with altered microbiota and increased susceptibility to certain intestinal pathogens. Indeed, gut LepRb cells are not epithelial but rather constitute a previously uncharacterized population of perivascular cells within the intestinal submucosa. Overall, our data reveal a role for LepRb signaling extrinsic to the intestinal epithelium and independent of food intake in the control of the gut microbiome. Dynamic interactions between gut microbes and the host modulate gut cellular proliferation, including the production of secretory cells and gut-associated immune cells (1). Dysregulation of the host-microbiome interaction may contribute to the pathogenesis of systemic metabolic disorders such as obesity (2, 3), metabolic syndrome (4), and cardiovascular disease (5). Shifts in fecal microbial populations correlate with obesity in both mice and humans, suggesting that certain bacterial constituents may modulate the susceptibility or response to weight gain (2, 6). One of the mechanisms by which gut bacteria may influence the host is through fermentation of otherwise indigestible dietary nutrients, rendering them available for host absorption, or through the generation of metabolites that modulate host biology (2, 7). Indeed, the absence of microbes in germ-free animals decreases caloric uptake from the diet and prevents diet-induced obesity. Whether obesity engages an adaptive mechanism to counteract chronic inflammation in adipose tissues has not been elucidated. Otop1 mutant mice respond to high-fat diet with pronounced insulin resistance and hepatic steatosis, accompanied by augmented adipose tissue inflammation. Thus, Otop1 defines a unique target of cytokine signaling that attenuates obesity-induced adipose tissue inflammation and plays an adaptive role in maintaining metabolic homeostasis in obesity. Obesity is associated with chronic low-grade inflammation in adipose tissues (59). The pathogenic role of the persistent activation of inflammatory signaling in metabolic disease has been demonstrated in numerous mouse models. An emerging view suggests that attenuating the proinflammatory response may provide significant metabolic benefits in obesity. Here we report that Clec16a is a membrane-associated endosomal protein that interacts with E3 ubiquitin ligase Nrdp1. Loss of Clec16a leads to an increase in the Nrdp1 target Parkin, a master regulator of mitophagy. Indeed, pancreatic Clec16a is required for normal glucose-stimulated insulin release. Thus, Clec16a controls b cell function and prevents diabetes by controlling mitophagy. This pathway could be targeted for prevention and control of diabetes and may extend to the pathogenesis of other Clec16a- and Parkinassociated diseases. Thus, it is critical to directly examine the functional role of potential disease genes and to correlate gene variation in potential enhancers to expression of the putative associated gene. Molecular understanding of new disease loci may provide important insights into the pathogenesis of human diseases and reveal new therapeutic targets (Pociot et al. Little is known of mammalian Clec16a function or of its role in disease pathogenesis. Here we discover a key role for Clec16a in the regulation of mitophagy, a selective form of autophagy necessary for mitochondrial quality control (Ashrafi and Schwarz, 2013). We find a key role for Clec16a in the maintenance of glucose homeostasis through its effect on the mitochondrial health of pancreatic b cells and, consequently, glucose-stimulated insulin secretion.