Low density lipoprotein (ldl) cholesterol - Free Essay Example

Sample details Pages: 17 Words: 5131 Downloads: 5 Date added: 2017/06/26 Category Health Essay Type Narrative essay Did you like this example? Elevated cholesterol, particularly low density lipoprotein (LDL) cholesterol, is a well defined risk factor for the development of atherosclerosis (NCEP, 2002). Atherosclerotic lesions are thought to form in response to endothelial damage caused by excess LDL (Steinberg, 2005). Most cases of cardiovascular disease (CVD), the leading cause of mortality and disability in the developed countries, are ascribed to pathology of atherosclerosis. Don’t waste time! Our writers will create an original "Low density lipoprotein (ldl) cholesterol" essay for you Create order Strategies to lower plasma LDL cholesterol levels are therefore crucial in the prevention and treatment of CVD (OKeefe et al., 2009). Conversely to plasma LDL levels, elevated high density lipoproteins (HDL) are protective against the development of CVD. Low density lipoproteins are responsible for the transport of cholesterol and fatty acids from the liver to the various tissues of the body. In contrast HDL is responsible for the majority of reverse cholesterol transport (RCT), in which cholesterol from the peripheral cells are returned to the liver for excretion in the bile (Joy and Hegele, 2008). It is this major role that HDL plays in RCT that is attributed with HDLs anti-atherogenic effects. Therefore strategies which elevate HDL levels are also beneficial in the prevention and treatment of CVD. Plasma cholesterol can be synthesized by the body (hepatic or extra-hepatic sources) or absorbed from the intestines (dietary or biliary sources). Statins, which are a family of HMG-C OA reductase inhibitors, reduce cholesterol synthesis and are very effective and can achieve plasma cholesterol lowering of up to 55% and have been shown to reduce CVD risk by one-third (Stein, 2002). Statin therapy has also been shown to increase plasma HDL levels from 2-16%, it is not know if this increase is clinically significant to the efficacy of statin therapy as it is difficult to separate this small and variable increase from the benefit of the significant LDL lowering (Hou and Goldberg, 2009). Regardless of the statins efficacy of return LDL cholesterol levels to their desired range, a significant proportion of statin users continue to have adverse events (Alsheikh-Ali et al., 2007). Therapies which reduce intestinal cholesterol or bile acid absorption are also available. Plant sterol/stanols, dietary fibre, bile acid sequestrants have been shown to be effective, with LDL cholesterol reductions of 10-15%, 8.5-13% and 5-30% respectively (Hou and Goldberg, 2009). The drug Ez etimibe, which binds to Niemann-Pick C1 Like 1 (NPC1L1) protein a crucial mediator of cholesterol absorption (Chang and Chang, 2008), has also been demonstrated to lower LDL cholesterol levels by 16-19% (Pandor et al., 2009). The wide ranges of cholesterol lowering seen in each treatment are likely the product of genetic factors which vary the rates of cholesterol synthesis and absorption, as well as modulate the effectiveness of each intervention. Because of this genetic heterogeneity of cholesterol synthesis and absorption, being able to effectively measure these parameters and how they respond to different dietary, pharmaceutical or lifestyle interventions is paramount to understanding interventions effect on cholesterol metabolism. Methods to assess cholesterol absorption Direct Method A direct method of measuring cholesterol absorption refers to measuring cholesterol flux from the intestines to the lymph. It is a direct method because it does not use a faecal or plasma level of cholesterol to infer absorption (Gibson, 1984). The first direct method of measuring cholesterol absorption required a duodenal cannula for infusion and cannulisation of the mesenteric or thoracic lymph ducts to sample lymph (Pouteau et al., 2003a). This method has been successfully used to access cholesterol absorption in rats, dogs and rabbits. The use of radioactively labelled cholesterol in the duodenal infusion allows for the distinction between exogenous and billiary cholesterol and can be used to calculate the percent absorption of exogenous cholesterol. An alternative direct method for use in humans was introduced by Grundy and Mok (1977). The intestinal perfusion method (sometimes refered to as Method VII) requires intubation with a three-lumen tube. Liquid formula and ÃÆ'Ã… ¸-s itosterol (though to be a non-absorbable cholesterol degradation marker at the time) is infused into the duodenum at the Ampulla of Vater though the first tube, and intestinal content is aspirated from the 2nd and 3rd tube, 10 and 100 cm distal to the first, respectively. Analysis of the aspirate from allows for the calculation of net hourly cholesterol absorption across the 100 cm long segment of intestine. Discriminating between exogenous and endogenous sources of cholesterol can be done by infusing radioactive cholesterol. The use of ÃÆ'Ã… ¸-sitosterol as a non-absorbable marker in this method may not have been the most appropriate choice given what is known about plant sterols and inhibition of cholesterol absorption at even very low levels (Calpe-Berdiel et al., 2009; Ostlund et al., 2002a; Ostlund et al., 2002b). Plant sterols complete with cholesterol for absorption into gut enterocytes, but are then exported back into the intestines by ABCG5 and ABCG8 transporters. This inh ibition was seen by Grundy and Mok, who noticed that even small increases in ÃÆ'Ã… ¸-sitosterol concentration caused noticeable reductions in measured cholesterol absorption (Grundy and Mok, 1977). The use of an inhibitor of cholesterol absorption in a method to access cholesterol absorption obviously leads to underestimation of actual cholesterol absorption capacity, and this must be taken into account when comparing absorption values among different methods. Another downside to the direct measurements techniques is their invasiveness, however they are the only methods which yield direct measures of total, exogenous and endogenous cholesterol absorption across a section of the intestine (Gibson, 1984). Cholesterol balance methods Cholesterol balance methods use the differences between dietary cholesterol and faecal cholesterol, excluding cholesterol of endogenous origins, to estimate cholesterol absorption (Matthan and Lichtenstein, 2004). Radioactive labelled cholesterol is used to distinguish endogenous from exogenous cholesterol and faecal and plasma levels of cholesterol are measured. There are 4 main balance methods, following the nomenclatures from Grundy and Arhens (1969), they are Method I, II, III and V. Method I consists of a single dose of [14C] or [3H] radio-isotope labelled cholesterol administered intravenously (Grundy and Ahrens, 1969; Quintao et al., 1971). This radio-isotope labels endogenous cholesterol and its subsequent steroid products. Using the formula: daily exogenous absorbed cholesterol (mg/day) = daily cholesterol intake (mg/day) à ¢Ã¢â€š ¬Ã¢â‚¬Å" daily unabsorbed dietary cholesterol (mg/day), and daily unabsorbed cholesterol (mg/day) = faecal neutral steroids per day (mg/day) endogenous faecal neutral steroids per day (mg/day). Endogenous faecal neutral steroids are calculated by dividing the total radioactivity (dpm/day) in the total faecal neutral steroids by the specific activity (dpm/mg) of the plasma cholesterol 1-2 days prior (depending on gastrointestinal transit time). Method II requires continuous oral labelling with radioactive cholesterol (Grundy and Ahrens, 1969; Quintao et al., 1971). The radio-labelled cholesterol is generally incorporated into a liquid diet to insure a precise daily intake of radiosterol for many weeks. Cholesterol absorption is calculated using the formula Z = X + Y, and Z * SA(Z)= X *SA(X) + Y * SA(Y), where Z = total faecal neutral steroids (mg/day), SA(Z) = specific activity of total faecal neutral steroid (dpm/mg), X = faecal neutral steroids of endogenous origin (mg/day), SA(X) = specific activity of plasma cholesterol (dpm/mg), Y= daily unabsorbed dietary cholesterol (mg/day), and SA(Y) = specific activity of diet ary cholesterol (dpm/mg). The equations are solved for Y, and absorption measurements can be done at any time after 4 days of continuous radioisotope feeding, a isotopic steady state is not required. It has been shown that Method II provides more consistent data on cholesterol absorption than, and is therefore recommended over, Method I (Gibson, 1984; Matthan and Lichtenstein, 2004). Method III was introduced by Wilson and Lindsey (1965) and requires an isotopic steady state. Continuous feeding with radioisotope labelled cholesterol for greater than 100 days is required to reach an isotopic steady state. When an isotopic steady state is reached daily cholesterol turnover is calculated, and daily absorbed dietary cholesterol is calculated by the formula: Dietary cholesterol absorption (mg/day) = daily cholesterol turnover (mg/day) * (specific activity of plasma cholesterol/specific activity of dietary cholesterol). Failure to reach an isotopic steady state will yield an underestim ate of actual dietary cholesterol absorption (Gibson, 1984). The long period required to reach an isotopic steady state and difficulty determining when this state has been reached make Method III a difficult and unreliable method. Method III was shown to give lower values of cholesterol absorption than both Method I and II in direct comparison (Quintao et al., 1971). Method V (Grundy et al., 1971) is a combination of methods I and II. Subjects are continuously fed isotopically labelled cholesterol as in Method I and then given a single bolus of different isotopically labelled cholesterol as in Method II. The equations for Method II are used, with only the modification of the X *SA(X) term which is replaced by [(R)(SA(X oral)) / (SA(X intravenous))], where R= daily faecal neutral steroid excretion of intravenous cholesterol (mg/day), SA(X oral) = plasma specific activity of oral isotope taken 1 day before R (dpm/mg), SA(X intravenous) = plasma specific activity of intravenous isot ope taken 1 day before R. Method V was designed for situation when rates of cholesterol synthesis are unusually high (due to surgical or pharmacological interference with enterohepatic circulation), which can lead to endogenous cholesterol being secreted into the intestines prior to isotopic equilibration(Gibson, 1984). In this circumstance (very high synthesis) Method I would underestimate and Method II would overestimate cholesterol absorption. Methods I, II and V all require a marker for gastrointestinal transit time, and for cholesterol degradation in the gut. Chromic oxide is often used as a marker of gastrointestinal transit time. ÃÆ'Ã… ¸-sitosterol has been used as a control for cholesterol degradation, however it must again be mentioned that if ÃÆ'Ã… ¸-sitosterol is used as a marker then the cholesterol absorption values obtained are likely lower than the actual values. However, in these methods if the degradation of cholesterol in the gut is not accounted for, the calc ulated cholesterol absorption will be higher than the actual values (Quintao et al., 1971). Isotope ratio methods Isotope ratio methods measure only the percentage cholesterol absorption (Pouteau et al., 2003a), unlike balance methods, and require accurate record of dietary cholesterol intake to estimate the mass of exogenous cholesterol absorbed from the percentage (Matthan and Lichtenstein, 2004). Method IV (also called faecal isotope ratio method), first introduced by Borgstrom (1969) calculates cholesterol absorption as the percentage of a single oral dose of radio-labelled cholesterol not recovered in the faeces. A single 1 ÃŽÂ ¼Ci radio-labelled dose of both cholesterol and ÃÆ'Ã… ¸-sitosterol is administered orally and feces are collected for seven to eight days. The ratio of labelled cholesterol to labelled ÃÆ'Ã… ¸-sitosterol in a sample of the pooled faeces samples is compared to the ratio that was given orally and cholesterol absorption is calculated using the formula: Cholesterol Absorption (%) = 1 à ¢Ã¢â€š ¬Ã¢â‚¬Å"((Faecal Cholesterol (dpm) /Faecal ÃÆ'Ã… ¸-sitosterol (dpm) ) * (Oral Cholesterol (dpm)/Oral ÃÆ'Ã… ¸-sitosterol (dpm))) *100. This method has been modified to allow a single faecal sampling, Sodhi et al. (1974) administered a faecal flow marker, such as chromic oxide or carmine red, with the original test dose, and calculated the faecal isotopic ratio at the peak level of the marker in the faeces rather than a sample of the pooled faeces. This single faecal sampling modification of Method IV has been compared to the original Method IV and Method I in baboons and it yielded consistently higher absorption levels, suggesting that this modification may weaken the accuracy (Mott et al., 1980). Method IV, despite the long period of faecal collection required, uses a far smaller dose of radioactivity than the balance methods, and is relatively straightforward and simplistic in execution (Gibson, 1984). It does still require ÃÆ'Ã… ¸-sitosterol as a marker of cholesterol degradation in the gut, so it shares the drawbacks previously discussed. Th e plasma isotope ratio method (IRM or Method VI), was first introduced by Zilversmit (1972) in rats and then subsequently carried out in humans (Samuel et al., 1978). The IRM involves simultaneous oral and intravenous administration of C14 and H3 radio-labelled cholesterol and requires only a single blood sample 3-4 days afterwards. The methods principles are based on the measurement of drug absorption used in pharmacology. If the absorption of cholesterol was 100% than the specific activity curve of both radio labelled cholesterols would be the same, if absorption was zero, none of the orally administered cholesterol, and therefore zero radioactivity from the oral radioisotope would appear in the plasma. Since cholesterol absorption falls between zero and 100%, the ratio of the two plasma radioactivities, after normalization for dose, are used to calculate absorption using the formula: Cholesterol Absorption (%) = 100 * (% oral dose in plasma/ % IV dose in plasma), where % oral dos e and % IV dose in plasma are the percentage of IV and oral tracer in the plasma sample, respectively (Samuel et al., 1982). The ratio is calculated 3-4 days after radioisotope administration because the specific activity time curves of both radioisotopes must be parallel. This does not occur in human until 3-4 days due to a delay in the appearance of the oral radioisotope in the blood related to the mechanisms of cholesterol absorption (Gibson, 1984). The IRM advantages over previous methods include: 1)only a single blood sample is required, 2) a low dose of radioisotope is used, 3)it does not require faecal collection 4) it does not required markers, such as ÃÆ'Ã… ¸-sitosterol, to correct for faecal losses. The IRM lends its self to repeated use because of its short duration and low level of labelling, this allows for investigation of variations in cholesterol absorption under different experimental parameters in the same individual in a comparatively short time period. The IRM m ethod has been validated in human numerous times against method IV under different conditions yielding similar results(Samuel et al., 1978; Samuel et al., 1982). A third isotope ratio method, Method VIII was introduced by Crouse and Grundy (1978), it shares similarities to Method IV, average cholesterol absorption is calculated using the ratio administered cholesterol to ÃÆ'Ã… ¸-sitosterol measured in the faeces, but differs in the method of isotope administration. C14 Cholesterol and H3 ÃÆ'Ã… ¸-sitosterol are administered orally three times daily for 10 days, and faeces can be collected from day 4-10 (Gibson, 1984; Matthan and Lichtenstein, 2004). Following day 3 of isotope administration the ratio of isotopes in the faeces becomes essentially constant and cholesterol absorption is calculated by the formula: Absorption (%)= 100* ((Faecal cholesterol (dpm)/Faecal sitosterol (dpm))/1-dietary cholesterol (dpm)/ dietary sitosterol (dpm)) This method requires only the ratio o f radioactivity in a single faecal sample to be measured; faecal mass need not be calculated (Crouse and Grundy, 1978). Since ÃÆ'Ã… ¸-sitosterol is also administered with the labelled cholesterol it may underestimate actual cholesterol absorption. Plasma cholesterol specific radioactivity following the consumption of a test meal has also been investigated a measure of cholesterol absorption. Lin et al. (2005) measured cholesterol absorption in 11 individuals with Smith-Lemli-Opitz syndrome (SLOS), a cholesterol synthesis disorder, and compared plasma cholesterol apecific activity with cholesterol absorption measured by Method IV. They sampled blood 24, and 48 hours following radioisotope enriched tests meals, and calculated the specific radioactivity of the cholesterol in the plasma (dpm/mg cholesterol). These values correlated significantly with cholesterol absorption calculated by Method IV (r=0.594, p=0.009, and r=0.474, p=0.047 for 24 and 48 hours respectively). While this m ethod cannot calculate cholesterol mass or percent absorption, it could allow for investigation of relative changes in cholesterol absorption within an individual across different conditions. It requires only a single blood sample, no stool collection, and no use of a cholesterol degradation marker, such as ÃŽÂ ²-sitosterol. This method is very similar, to the single isotope tracer method that will be discussed in the stable isotope methods section (Wang et al., 2004). This relationship between plasma radioactivity at 24 hours and cholesterol absorption requires further validation, especially in healthy individuals. The use of radio-isotopes has been invaluable to the investigation of cholesterol absorption, however the advent of stable isotope laboratory techniques, especially developments in isotope ratio mass spectrometry has allowed radio-isotopes such as C14 and H3 tracers to be replaced with safer 13C, 2H and 18O stable isotope labelled tracers (Pouteau et al., 2003a). T his has reduced the difficulty related to containment, handling, disposal and overall safety associated with radioisotopes and allowed for investigation of cholesterol absorption in certain populations (children, pregnant and lactating mothers) which had previously been impossible do to the ethical considerations around radioisotope administration. Stable isotope methods for cholesterol absorption Cholesterol labelled with stable isotopes has been shown to have identical kinetics as radio-labelled cholesterol. This has led to the development of stable isotope techniques to investigate cholesterol absorption. Lutjohann et al. (1993) introduced a stable isotope version of Crouse and Grundy s (1978) Method VIII discussed above. Deuterated cholesterol and sitostanol were used in place of C14 cholesterol and H3 ÃÆ'Ã… ¸-sitosterol, and quantified using gas-liquid chromatography -selected ion monitoring. Cholesterol absorption was calculated as in Method VIII. The stable isotope method was twice compared to Method VIII, in six monkeys, yielding similar results. The stable isotope method produced and absorption range of 49-73% (mean of 60%), with coefficient of variation ranging from 3.9%-15.1% (mean 7.1%). The radioisotope produced a range of 51-69% (mean 61%) with coefficient of variation ranging from 1.9-13.6% (mean 5.1%) (Lutjohann et al., 1993). This stable isotope Method VIII was determined to be as effective as the radio-isotope Method VIII, without the risk of radioactive exposure to subjects and research staff. This method uses sitostanol, rather than ÃŽÂ ²-sitosterol, as a marker for faecal losses of cholesterol. This, however, does not remedy the problem other methods suffer from when using ÃŽÂ ²-sitosterol, as sitostanol is also an inhibitor of cholesterol absorption (Gylling et al., 1997; Miettinen et al., 1995). The plasma isotope ratio method (Method VI) (Samuel et al., 1982; Zilversmit, 1972) was also adapted to stable isotopes. Bosner et al. (1993) used 2H labelled oral and 13C labelled IV cholesterol to calculate the plasma stable isotope ratio and cholesterol absorption percentage. This method uses gas-chromatography à ¢Ã¢â€š ¬Ã¢â‚¬Å" mass spectrometry with select ion monitoring (GC/MS-SIM) or GS/MS- chemical ionization mode (CI) to determine isotopic enrichment. Bosner et al. (1999) further modified the Method VI to a single isot ope dual tracer method, using oral [2H]5 and IV [2H]6 cholesterol. Isotope detection in the plasma cholesterol is done by GC/MS- selected mass monitoring. Jones et al. (2000) were the first to use isotope ratio mass spectrometry to determine isotopic enrichment using the plasma isotope ratio method. Oral 13C and IV 3H cholesterol were administered to 15 hypercholesterolemic men, followed by blood sampling 2-3 days post administration. Free cholesterol from red blood cells was purified by thin layer chromatography and subsequently combusted to yield carbon dioxide and water. The CO2 was then measured for 13C enrichment against the international standard Pee Dee Belemetite (PDB) on an isotope ratio mass spectrometer (IRMS). The water was reduced to hydrogen gas via zinc reduction and 3H enrichment was measured against Standard Mean Ocean Water (SMOW) international standard by IRMS. The ratio of plasma enrichment of 13C to 3H cholesterol on day 3 after tracer administration was used to calculate cholesterol absorption. Recently, continuous flow gas chromatograph pyrolysis IRMS (GC/P/IRMS) systems and 18O or 3H cholesterol with gas chromatograph combustion IRMS (GC/C/IRMS) systems and 13C cholesterol have been used in calculating cholesterol absorption using IRM (Method VI)(Gremaud et al., 2001; Pouteau et al., 2003b). The use of IRMS vs GC/MS-SIM has vastly increased the precision of this method. The two single stable isotope methods for accessing cholesterol absorption were introduced by Ostlund et al. (1999) and Wang et al. (Wang et al., 2004). Ostlund et al. (1999) administered deuterated cholesterol to volunteers and measured the average oral cholesterol tracer in plasma ((mmol deuterated cholesterol/mol natural cholesterol) in blood samples taken 4 and 5 days post tracer administration using GC/MS. Wang et al. (2004) administered 13C cholesterol orally followed by blood sampling at 24, 48, 72 and 96 hours. 13C enrichment in plasma free cholesterol was mea sured using GC/C/IRMS. Average 13C enrichment from 24-96 hours and area under the curve (24-96 hours) of 13C enrichment were compared the cholesterol absorption percentage measured using the plasma dual stable isotope ratio method (Bosner et al., 1993) in 2 studies. The average and area under the curve of 13C enrichment in plasma free cholesterol correlated with cholesterol absorption percentage measured by stable isotope method VI (r values ranging from r=0.81, p=0.0001 to r=0.88, p=0.0001)(Wang et al., 2004). Both of these single isotope methods are used to compare treatment effects, such as pharmaceutical or dietary interventions and their effects relative to control, on cholesterol absorption. Absorption surrogate method The use of serum plant sterol levels to predict cholesterol absorption was first developed by Tilvis and Miettinen (1986). They showed that serum levels of ÃÆ'Ã… ¸-sitosterol and campesterol, when normalized for total serum cholesterol, correlated positively with cholesterol absorption as measured by Method VIII of Crouse and Grundy (1978). This measurement involves lipid extraction from a single blood sample followed by GC or HPLC to quantify serum plant sterol and cholesterol levels. The use of campesterol or ÃÆ'Ã… ¸-sitosterol to cholesterol ratio has since been used numerous times as a measure of cholesterol absorption (Gylling et al., 2007; Hallikainen et al., 2006; Matthan et al., 2009; Nissinen et al., 2008; Simonen et al., 2008). Nissinen et al. (2008) showed that ÃÆ'Ã… ¸-sitosterol to cholesterol ratio correlated better with cholesterol absorption than campesterol to cholesterol ratio, as measured by method VIII, across three diets varying in both cholesterol and fat le vels in 29 healthy male volunteers. It is important that when using serum plant sterol as surrogates for cholesterol absorption it is imperative that factors which are known to change serum plant sterol levels, such as the dietary intake of plant sterols (Chan et al., 2006), should be controlled for so as not to perceive a change in cholesterol absorption which may not exist (Vanstone and Jones, 2004). Serum plant sterol levels can be variable within and across different population (Chan et al., 2006), they can also be severely elevated in certain individuals due to genetic disorders (Berge et al., 2000; Lee et al., 2001). It is important that individuals with these genetic disorders are not included in studies using this method of estimating cholesterol absorption as serum plant sterol levels do not reflect cholesterol absorption in these individuals. Recently it has also been shown that serum plant sterol concentrations do not reflect cholesterol absorption in individuals with Smi th-Lemli-Opitz syndrome (Merkens et al., 2009), therefore the use of serum plant sterols as surrogates for cholesterol absorption should be verified prior to its use in a particular populations. The use of plant sterol surrogates does benefit from relative speed and simplicity compared to other previous discussed methods (Matthan and Lichtenstein, 2004; Pouteau et al., 2003a), it is the only estimate of cholesterol absorption which can be done in large scale studies. Methods to access cholesterol synthesis The primary pharmaceutical intervention used to lower elevated cholesterol levels are statins, which are inhibitors of HMG-CoA reductase, a key enzyme in cholesterol synthesis (Stein, 2002). Cholesterol synthesis contributes more to circulating cholesterol levels than cholesterol absorption (Dietschy, 1984; Pouteau et al., 2003a), and maintains cholesterol levels during fasting, therefore accurate assessment of cholesterol synthesis is essential to the field of cholesterol research. Balance Method Cholesterol synthesis can be estimated when intake of dietary cholesterol and excretion of total cholesterol is known during a metabolic steady state (Grundy and Ahrens, 1969). The criteria of this metabolic steady state are: constant plasma cholesterol and faecal cholesterol excretion levels during a period of constant weight. In this steady state, cholesterol synthesis is the difference between cholesterol excretion (faecal neutral sterols and bile acids) and intake. Dietary cholesterol intake must be accurately measured, and faeces must be collected, for a given period of time. Faecal flow must also me monitored to assure collection of faeces from the appropriate time period. Faecal neutral sterols and bile acids are measured in the faeces, typically by GC-MS, and cholesterol synthesis for a given period of time is calculated (Jones et al., 1998; Kempen et al., 1988). Although this method is the gold standard for calculating cholesterol synthesis, it is vulnerable to errors in t he estimation of both cholesterol intake and excretion which can potentially cause significant error. The balance method does determine the actual mass of cholesterol synthesized during a given period, it requires a metabolic steady state, accurate measurement of cholesterol intake and total faecal collection for the period of interest. Therefore this method is not suited for larger trials. Fractional conversion of squalene Daily cholesterol synthesis rate can be estimated by the fraction of infused radiolabelled melavonic acid converted to cholesterol (Liu et al., 1975; McNamara et al., 1977). [14C] mevalonic acid and [3H] cholesterol are administered intravenously, this infusion of labelled mevalonic acid rapidly labels the plasma squalene pool, reaching a maximum enrichment at ~100 minutes. Cholesterol synthesis is estimated by measuring squalene synthesis. Squalene synthesis is calculated by the percentage of melavonate dose converted to cholesterol, divided by the area under the curve of pasma squalene specific activity. This method assumes that plasma squalene synthesis is equivalent to cholesterol synthesis. The cholesterol synthesis rate estimated using this method has been shown to agree with cholesterol synthesis calculated by the balance method within 8% (Liu et al., 1975). The benefits of this method is it requires only 1 hour of a participants time, and can be repeated every 3 weeks (McNa mara et al., 1977), it does however require the administration of intavenous radioisotopes. Cholesterol synthesis precursors The concentrations of plasma precursors along the synthesis pathway of cholesterol have been used to as an indirect qualitative measure of cholesterol synthesis. Squalene (Miettinen, 1982), mevalonic acid (Parker et al., 1982), lanosterol, desmosterol and lathosterol have all been used a surrogates for cholesterol synthesis (Matthan et al., 2000; Miettinen et al., 1990). These precursors have has been shown to fluctuate with diurnal cholesterol synthesis patterns as well as in conditions in which cholesterol synthesis is elevated or reduced (Miettinen, 1982). These precursors correlate more closely with measured cholesterol synthesis when they are normalized for plasma cholesterol level, then as the absolute amount of precursor, and are normally expressed in mmol/mol cholesterol (Miettinen et al., 1990; Nissinen et al., 2008). When using cholesterol precursors as surrogates for cholesterol synthesis, dietary intakes of each precursor should be controlled for, this is particularly i mportant for squalene, which is abundant in olive oil and is at least partially absorbed into the blood (Ostlund et al., 2002b). The most consistent surrogate for cholesterol synthesis has been lathosterol (Kempen et al., 1988; Nissinen et al., 2008). Since cholesterol synthesis surrogates require only one blood sample they are ideal for estimating cholesterol synthesis in very large studies or epidemiological trials. Mass Isotopomer Distribution Analysis (MIDA) Mass isotopomer distribution analysis (MIDA) is a technique that can be used to measure the synthesis of biological polymers in vivo (Hellerstein and Neese, 1999). The technique uses the relative abundance (pattern or distribution) of polymer species which differ only in mass (mass isotopomers) produced during the administration of stable isotope labelled precursors. The distribution of the polymer species produced is compared to the theoretical distributions predicted by binomial and polynomial expansion. Using these theoretical distributions, parameters such a fractional synthesis rate can be calculated using combinatorial probability model. Since cholesterol is synthesized from subunits of acetyl-Coa, fractional synthesis of cholesterol can be calculated during the infusion of 13C labelled acetate (Neese et al., 1993). This method is very invasive, requiring a 24-hr intravenous infusion and serial blood sampling from a indwelling catheter (Di Buono et al., 2000). The cholesterol in the blood samples is measured by GC-MS to determine the distribution pattern of isotopomers from which the rate of synthesis is obtained. The data analysis relies on more complex mathematical modelling than other methods for estimating cholesterol synthesis. Deuterium Incorporation This method is based on the tritiated water uptake method by Dietchy and Spady (1984) used originally in animals. Deuterium incorporation method uses water as a tracer to determine the synthesis of free cholesterol (FC). The fractional synthesis rate (in pools/day) of free cholesterol is calculated from the rate of incorporation of deuterated water into de novo synthesized plasma or erythrocyte cholesterol. Orally administered deuterated water equilibrates with the bodys water and NADPH. Body water and NADPH account are the precursors for 22 of the 46 hydrogen in synthesized cholesterol (Jones, 1990). Deuterium enrichment of the precursor pool, plasma water, and in erythrocyte or plasma cholesterol are measured by IRMS. Fractional synthesis rate of free cholesterol (FSR-FC) is calculated using the following formula: FSR-FC (pools/day) = ( ÃŽÂ ´ cholesterol à ¢Ã¢â€š ¬Ã‚ °/ ÃŽÂ ´ plasma water à ¢Ã¢â€š ¬Ã‚ ° * 0.478) ÃŽÂ ´ refers to the change in deuterium enrichment over 24 hours and 0.478 is the ratio of cholesterol from body water and NADPH to total hydrogen in a cholesterol molecule, or the ratio of hydrogen which could be enriched by oral D2O administration (Jones et al., 1993). From the FSR-FC the ASR -FC (g/day) can be calculated by multiplying the FSR-FC by the M1 pool size (determined by Goodmans equation) and 0.33 the proportion of FC in total cholesterol. The ASR-FC is approximately the daily production of newly synthesized cholesterol. The three main assumptions required for deuterium incorporation are: 1)That the fraction of hydrogen derived from plasma water (22/46, or 0.478) is constant in denovo synthesized cholesterol. 2) That denovo synthesized free cholesterol rapidly exchanges between the site of synthesis and the major free cholesterol (plasma compartment) pool. Within this pool cholesterol migrates rapidly between cellular membranes and lipoproteins as well as between different classes of lipoproteins. 3) Deuterium uptake i nto the free cholesterol of the major (plasma) pool of cholesterol reflects synthesis in this pool and that synthesis of cholesterol in the major pool provides a reasonable measurement of total cholesterol synthesis as most sterol synthesis occurs in the intestines and liver which contribute to the plasma cholesterol pool. It must be acknowledged that the major (plasma) pool of cholesterol is at equilibrium with two other slow turnover pools outside the plasma, and that the slow inter-pool cholesterol exchange would cause insignificant entry of labelled free cholesterol into the central pool within a 24 hour time period (Dietschy and Spady, 1984; Jones, 1990). Although it has been shown that these assumptions are not perfect, the cholesterol synthesis estimates yielded by deuterium incorporation have been sensitive enough to show differences in cholesterol synthesis due to genetic factors, and dietary and pharmaceutical interventions. It has also been shown to correlate well with ch olesterol synthesis measured by the balance method (Jones et al., 1998), MIDA(Di Buono et al., 2000) and cholesterol synthesis surrogate levels (Matthan et al., 2000). Conclusion The methods used to quantify cholesterol absorption and synthesis reviewed in this article have yielded invaluable information, as well as provided effective means of measuring the experimental effects of various interventions, on cholesterol homeostasis. The methods have evolved substantially over the years, from radio-isotopes to stable isotopes, and from highly invasive to less invasive procedures. However, advancement in the quantification of cholesterol homeostasis must still be continued, with a goal of finding more accurate methods to measure cholesterol synthesis and absorption, perhaps simultaneously, something which until now is still unavailable. The advantages and drawbacks, as well as the type of information yielded by each method for measuring cholesterol absorption and synthesis must be weighed carefully when selecting an appropriate method. The cost and available technical expertise and facilities will limit which methods are available to each investigator. However, it is imperative that the assumptions and limitations of each method are checked to insure that its use is valid for each particular experimental question it is used to answer.

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The damage and loss of life has caused significant delays to the release of LPHGs release of EFHG. DRP Documents: (e.g., disaster recovery plan, backup plans etcRead MoreSteps for Conducting a Postmortem1523 Words   |  6 PagesConducting Post-Mortem The following are the steps taken for conducting an incident post mortem; first, is to get a clear picture of the state of affairs. It is critical to analyze network maps, talk to the key witnesses of the disaster, learning the occurrences from the data available from the weather department in a bid to collect evidence. At this stage getting every log available is useful, thereby casting a wide net in order to capture the details of the occurrences including the causes.Read MoreRecovery Options : Disaster Recovery Plan1186 Words   |  5 PagesRECOVERY OPTIONS Disaster recovery plan consists of protecting and recovering information systems to resume normal business operations in the most cost-effective manner. 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The Management Efficiency Unit Free Essays

I have been working in the Public Registry for the last 8 years.   The location of the offices is quite distant from my residence and sixteen other colleagues that work in the aforesaid department have the same problem.   Every morning we are required to travel a one hour ferry and nearly 2 hours drive to arrive at work. We will write a custom essay sample on The Management Efficiency Unit or any similar topic only for you Order Now The Management Efficiency Unit noted this issue and finally top management decide to take remedial action by requesting a transfer to another department, the Treasury Section, which mitigates such location problem. Unfortunately the scheme designed by executive management was not properly planned.   They also did not discuss this issue with operational managers who possess far greater knowledge on the day to day running of the office.   As a result the decision lacked practical issues and led to other problems, as will be further illustrated below. Everybody accepted the transfer and we were eventually informed that our training for the new job will take approximately two or three months and will take place in the head office of the Finance Department, which is also distant.   The new job was highly technical and involved a lot of accountancy issues, which the majority of us were not capable to comprehend that easily. The three months passed and we were still unable to work unsupervised from our tutors.   The impracticality of the decision was highlighted from the fact that our principal in charge, who was also taking training was still incapable to work on his own after three months. When these issues started to crop up, crises management arose.   A new assistant director was assigned to this division, who once realized of the vast serious problems arising from such decision, he immediately denied this new role. Demotivation arose in our section and staff is constantly complaining about the problems we are facing.   In addition no communication exists between us and executive management and we are alone with this new work, which we are not trained to do. The only reasonable solution that management was able to come up with is to extend the duration of the training to eight months.   However with this decision they have not yet solve the problem that the location of the office is far from home and are incurring additional costs to train us. If they planned carefully before deciding and discussed it with operational managers they would have definitely solved the problem more efficiently.   It is imperative that when managing change proper communication and planning exist. Reference: Johnson G.; Scholes K.; Whittington R. (2005). Exploring Corporate Strategy. Seventh Edition. England: Pearson Education Limited.    How to cite The Management Efficiency Unit, Essays

China Social Structure free essay sample

It will outline the different class’s that make up contemporary China’s social structure and give a detailed outlook and perspective on each class, and show what change they have undergone since the opening of China’s economy in the late 1970’s and introduction to a market based economy. The greatest outcome will see how the transfer of the class’s from a socialist dictated economy and society during the Mao era, rapidly changed and fused into the modern market based economy of today’s China. This essay should also indict who has benefited most from such a quick and bold move to a market economy, and those who have lost out and not been so lucky as others due to the open door policy of China which was introduced in 1978, by then Chinese Premier Deng Xiao Ping ( ). This essay will take each class individually and contrast them to other class’s, both those that existed during the Maoist era of pre-1978 and the class’s that have emerged as a result of the economic reforms pursued by China since the opening of its economy and internal reforms where introduced. We will write a custom essay sample on China Social Structure or any similar topic specifically for you Do Not WasteYour Time HIRE WRITER Only 13.90 / page Lastly it will look at if China’s communist party has steered away from the founding ethics of a socialist economy to that of a capitalist one due to social class division and what effect this can have on China in the near distant future. Firstly looking at the Peasant class, one of the three original social class’s during the Maoist period of 1949-1978, (the other two being the working class and the cadre class). The peasant class, along with the working class during Maoist China were dubbed the proletariat class, in comparison the relatively small but evident cadre class. The rural-urban divide has always been existent in Chinese society, largely based on economic and geographical contributions. However throughout the Maoist era, peasant’s standard of living; to a certain extent were raised. With the abolishment of savage landlords which persisted during the imperial and republican times and the introduction of many yet simple beneficiaries to rural areas of China, peasant’s standard of living from 1949-1976, actually rose significantly, ‘On the one hand, standards of living can be seen as improved due to the absence of warlords, bandits, landlord, and local tyrants. The government invested a lot in agriculture, especially water conservancy, irrigation works, chemical fertilizers, and agricultural machinery†¦. The life expectancy of peasants increased from less than forty years before 1949 to more than sixty years in the 1970s’[1] Simple improvements in education and health, the fundamentals of any society to prosper were drastically improved by the so called â€Å"barefoot† teachers and doctors. However, prosperity was limited due to collectivisation of all the land available for agriculture, restricting peasant’s income and also led to the disaster of the Great Leap Forward campaign, which saw many peasants suffer in comparison to their urban counterparts. Restrictive movements of people from rural to urban also led to a stagnated peasant society, with the introduction of the household registration system ( ) , ensuring that peasants and their families never had the chance to seek a more prosperous life in the cities or enjoy the benefits of those of urban household with hukou registration. The reforms of 1978 first and profoundly had an immediate effect on rural areas. This came with the abolishment of the commune system, establishment of free market practice in the countryside with agricultural products and the thriving success of the Town and Village Enterprises (TVE’s). By 1993, 145 million peasants had become members of the working class; however they would be referred to as peasant worker, and not urban worker due to the hukou registration system. (Li Yi, China Startifictaion, p. 105). Many rural inhabitants, after the reform era had taken jobs in cities, albeit as a floating population, due to the strictness of the hukou household registration system most rural registered workers in cities today don’t enjoy the benefits of their urban registered counterparts, of housing, health care and schooling for their children. Most rural areas are also responsible for development of their own areas, with little support from the government, taxes and fees remain high in proportion in the countryside, as is the cost and low opportunities of schooling and further education. Li Yi China Stratification p. 192)Peasants also missed out largely in China’s great economic boom during the 1990’s and early 2000’s, with China’s economic annual growth an average of 9% in recent years, peasant income has increased, but rather in some cases decreased (Li Yi China Stratification p. 219). Peasants are in a sense stuck within their boundaries due to the House hold Registration System, or hukou. This social â€Å"apartheid† has created a massive imbalance between the coastal urban areas and the mainly rural western half of China. Urban household registered hukou holders had much of the states benefits in housing, education healthcare and employment ( Fei Ling Wang, Chinese Society, Change, conflict and resistance, (New York, Routledge, 2000). But as with the opening reforms of the late 1970’s, not all urban households and workers have managed to benefit, with that creating class differences even within China’s urban populace. Throughout the Maoist era, most urban employment was contingent within State Owned Enterprises(SOE’s), with workers benefiting from the system dubbed the â€Å"iron rice bowl†. This system, commonly a factor of socialist geared economies and societies was the benefits urban state employees enjoyed throughout Mao’s reign. Benefits included housing, food rations, healthcare insurance and education of employee’s children. The â€Å"iron rice bowl† also supplied life long work and benefits to those who were employed with work units, danwei ( ) and simplified that throughout the socialist market era, little social conflicts and imbalanced occurred amongst the urban working class. As of 1978, as much of 78% of urban labour force were recipients of this â€Å"iron rice bowl† welfare. [2] Starting with the reforms initiated by Deng Xiao Ping, slowly the â€Å"iron rice bowl† was withdrawn and open competitiveness was encouraged. Seen as extremely costly to the state, SOE’s would now undergo huge transformation. â€Å"Grasp the big, release the small† was now in effect, with SOE’s that were making a loss to face closure, and the ones that remained facing intense competition from the more effectively run foreign enterprises and privately run sector. This in effect led to high levels of lay offs, never heard or seen of during the Mao regime, with estimates as high as 60 million, with an overall figure including those waiting on employment reaching 100million. [3] The huge amount of layoffs had never been experienced in China, and the sheer pressure on the state took hold, not all of employees who had lost their â€Å"iron rice bowl† could find work, due to the fact, they had worked in industries all their lives that largely included the same repetitive work, and had no grasp of competitiveness. Much of the lay offs were women, and above average age workers, the category that will find it hardest to find re employment. Much lacked a good education, having experienced the chaos of the cultural revolution of 1966-1976, which saw China’s education, especially upper levels deteriorate. This urban class, dubbed the xiagong, literally went from complete stableness in the â€Å"iron rice bowl† system to a complete foundation less society. Alongside the rural migrants of the â€Å"floating population†, who’s number in cities is rising, coincided with the now jobless former state urban workers. Competition for employment was extensive. With a poor social security service, or none at all, these former secure workers now faced a much tougher life to secure an income to support themselves and families, with wages below the average of urban household incomes, ‘it is not surprising that by early 2000, 73 per cent of China’s urban population had incomes below the national average, and just 27 per cent were above it,’[4] A real sense of anxiety surely crossed the minds of those who lost out, having gone from a society that provided everything to one now having to defend for themselves with the handicap of age, lack of education and skills that were needed to fill the positions now open to the market economy. Not only did they see there jobs and way of lives disappear, state benefits also dried up, or were not enough to help support this group of former state workers. The numbers who fell into poverty, which is classified depending on which city it is in China reached startling highs of almost 13 per ce nt of urban population, 40-50million, [5] in 2001. The unemployment figures, relating from state owned enterprises and cooperatives seems to have increased as of 2001, with China’s acceptance into the World Trade Organisation (WTO). The state-society model, developed throughout the Maoist years seems threaten, with the massive layoffs of these ordinary workers, however much of the managerial staff and cadre’s who where involved in the running of these SOE’s have benefited, becoming prime shareholders, re allocated to improved positions of employment and benefits that go with that, again dividing urban class standards, ‘People working in rich work units could easily get a comfortable spacious apartment, while those in poor work units remained in near-slum conditions. Work units ability to provide housing varied between state and collective sectors and with bureaucratic rank. While work unit housing was allocated to satisfy needs (large or multigenerational families were allocated first and got more living space) spacious and quality units were a work units resources and served as incentives to reward political and managerial authority, seniority , professional expertise, and social connections . ’[6] At the expense of the ordinary workers losing their occupations in the numbers, a new class of highly educated, well positioned (through connections like guanxi ( ) and entrepreneurial class has reaped the real rewards of the booming Chinese economy of the recent years. Have the government and the ruling Communist Party really shown nepotism away from its own founding base of the working class? What can the workers do t voice their concerns, in a country that restricts free speech and emotion? Any organisations of protests are met with harsh retaliations, and those accused of such activities are harshly dealt with. Taking the example of the Daqing petroleum protest of March 2002, in Daqing Heilongjiang province, the state must address and deal with these challenges sensitively, knowing that a lot rests on its outcome. In dealing with large scale outbreaks of protest like this the state has developed a ‘divide-and-rule strategy, encouraging employed workers to keep their distance from the protests, and intentionally causing the rank and file among the unemployed to believe that the protests are to blame for the cancellation of payments’[7]. This form of catch-22 policy has so far deemed successful in maintaining stability and control over any protest or arguments amongst the newly found unemployed â€Å"iron rice bowl† employees. This has allowed the government to keep on track with its promotion of the market based economy on a whole, as well formulating a large labour surplus into the workforce. The urban working class has definitely been the class that has lost most in the post-reform era of China, while a new class has benefited at their downfall. Reform era China has provided vast opportunities in newly emerging industries and privately owned, foreign invested or created from scratch enterprises have thrived. However demand for educated, well positioned workers, has left modern China with a development of a â€Å"middle class†. It is hard to say what exact characteristics make up this â€Å"middle class†, as in comparison to western middle class society, it is far from a following example, like the ownership of cars as common in western income families in the west is obviously not matched in China. In terms of income, an annual household income of 60,000-500,000 yuan is thought to qualify a household as middle strata levels. 8] This newly educated class, with social guanxi is the new driving force of China’s growth, with the closing of SOE’s and rising capitalist activity in the Chinese economy. Newly developed entrepreneurs, officially welcome into the Communist Party in 2001, by invite of the Thr ee Represents, have been behind much of China’s economic activity, able through connections, know how, and close connections to the party(Goodman, New Rich in China, p34-36) been able to secure funds to help develop their prestige. Professionals and managers have also been on ends of high salary turnovers, seen to the state as vital in its strategic to immense economic activity. Favours from the state also passed their way through to this new elite group, with property and housing given at lower rates, ‘the massive sale of public housing to employees throughout the 1990s occurred at highly subsidized prices for the existing housing stock, or alternatively employees were given the option of buying newly built houses while the work unit carried the lion’s share in construction or purchasing costs†¦buying extensively to cater for the needs of their professionals and other employees. ’ [9] These new homes ‘awarded’ to the newly emerging elites are often found close to the best schools and other community services, creating so called neighbourhood apartheid. An estimated 20-30 billion yuan was lost due to under value sales of land, ( www. internationalviewpoint. org/spip. php? article751) . So why has the state in turn favoured the class that had been, during the Maoist era received massive vocal attacks? Well in dealing with the massive under performing SOEs , the state could and did look to the emerging capitalists as a source of absorbing the loose employment that spilled out of the massive state owned unemployed. Capitalists are thought to have accounted for between 70-85% of China’s GDP ( Li Yi, China Stratification, p137), without this China may well have internally collapsed, unwilling to disband its loss making enterprises and not indulging into a market economy. This system, albeit seeming unfair, is following in Deng Xiao Ping’s approach of allowing some people to get rich first’. However this route, of eventually reaching a society where the middle class is prominent and thriving will take time and the correct policies and approach, with careful management.

Gas War Essays (2040 words) - Petroleum Politics, Commodity Markets

Gas War Gasoline is produced by a distillation process where crude oil is heated and fumes are captured and converted into many products such as kerosene, jet fuel, and gasoline to name a few. Therefore the price of crude oil, which is extracted from oil wells beneath the earths surface, is a major factor in gas prices. The five leading oil-producing countries and their approximate shares of the world supply of oil are: Soviet Union 21%, Saudi Arabia 17%, The United States 15%, Venezuela 4%, and Mexico 4%. These five countries made up 61 % of the worlds oil production back in 1980. Even though The United States is a major producer of oil, it does not make them self-sufficient. The United States uses more oil than they can produce and must look towards foreign countries. An organization called O.P.E.C. controls approximately four fifths of the worlds oil reserves in the non-communist world. The United States is forced to deal with O.P.E.C., not only in its own interests, but also in the inter est of its allies and in the interest of maintaining peace. The former Soviet Union may now have an interest in selling some of their oil that they have a tremendous amount of. O.P.E.C. which stands for Organization of Petroleum Exporting Countries, is made up of 13 countries: Iran, Iraq, Kuwait, Saudi Arabia, Venezuela, Qatar, Indonesia, Libya, United Arab Emirates, Algeria, Nigeria, Ecuador, and Gabon. O.P.E.C. was founded in Baghdad, Iraq in September of 1960. It was organized in response to oil producing countries that did not consult with the Middle Eastern oil states before lowering their crude oil prices. The producers feared that other countries would establish monopolies. The aim of O.P.E.C. was to create a universal price between the countries, in order to ensure peace between oil producers throughout the world. O.P.E.C. also wanted to provide its members with technical and economic support in times of need, since not all the countries were completely stable. The headquart ers were initially set in Geneva, but were later moved to Vienna in 1965. O.P.E.C.'s goal was to establish firmly unified prices amongst their members, but the organization was not always successful. In their quest for control over the world market of oil production, they have ran into several obstacles and setbacks. O.P.E.C. has barely survived being eliminated due to internal conflicts amongst its members. Since O.P.E.C. almost has a strangle hold on the worlds oil supply, The United States is extremely concerned with the areas instability. The Middle East and the Persian Gulf area, where most of the members are located, are extremely prone to wars, both civil and cross borders, plagued by religious battles, and positions of power are frequently overthrown, making it hard for any stability to come out of the area. Any time there is chaos in the Middle East, The United States thinks back on ?memories of other troubles in the Persian Gulf area: the Arab oil embargo in 1973-74, the Iranian revolution in 1979-80 and Saddam Hussein's invasion of Kuwait in 1990. (1) The area is also vital to our allies, who would be crippled without Gulf oil, whose livelihood we are dependent on. In 1973 O.P.E.C. raised oil prices 70%. The dominant Middle Eastern members of O.P.E.C. used succeeding price increases as a political weapon aimed at Western nations in retaliation for their support of Israel against its Arab neighbors in the so-called Yom Kippur War of October 1973. Prices were accordingly raised another 130% at the Tehran conference of December 1973, and a temporary embargo was placed on the United States and the Netherlands at the same time. Other prices increases followed in 1975, 1977, 1979, and 1980, which ultimately raised the price of a barrel of crude oil from United States $3.00 in 1973 to $30.00 in 1980. (2) Almost every college student has heard stories from friends or relatives about the gas crunch in the 1970's. People waited in lines that stretched for miles, and could only get gas on certain days depending on the first letter of your last name. O.P.E.C. used the money they raised to invest in other countries, placed in foreign banks, currency markets, and to help their

Cinema is an Adventure , but the difficulty is that it is an adventure that you have to strive continually to control The WritePass Journal

Cinema is an Adventure , but the difficulty is that it is an adventure that you have to strive continually to control Abstract Cinema is an Adventure , but the difficulty is that it is an adventure that you have to strive continually to control AbstractIntroductionJean Rouch as a Film maker and an Ethnographerâ€Å"Les Maitres Fous† (The Mad Masters)ConclusionBibliographyRelated Abstract The very perceptions and style of ethnographic cinema was changed with the intervention of late Jean Rouch’s films. He was considered as one of the founding fathers of Cinema Verite   or Real Cinema In France,   and Surrealism. Jacques Rivette (1968) considers Rouch as the force behind the entire French cinema made in past ten years. Rivette comments that the importance of Rouch is much more in the evolution of French Cinema than Godard with few people realizing it (Rivette,1968). According to Henley(2009) his sudden death in a road accident, in February 2004, made Visual Anthropology lose one of its precious gems. Rouch was a visionary and a true believer in the strength of lenses. In most of his interviews he talked about the power of camera and the responsibility attached to it. He believed that camera had the power to reveal the inner most beliefs and impressions of mankind (Henley,2009). My first encounter with Rouch’s films was in my Visual Anthropology classe s. At first I found it quite difficult to understand his films as most of them were in French. However Rouch’s different and controversial approach to ethnographic cinema with his very interesting personal touch in all his films molded me to watch his movies in spite of such difficulties. Henley comments that though Rouch made more than one hundreds of films but only handful films have been distributed far wide while majority of the films remain unknown and difficult to see specially for English speaking world (Henley,2009). The most interesting fact about Jean Rouch’s cinema, as Mick Eaton puts forward was that, Rouch believed that more observations can be done with a film rather in direct observations. He suggested that through films you can get more feedback from your subjects and participants which make the observation far more interesting and trustworthy. Rouch belived that his films were for greater number of people, it is a fight against the colonial oppressions and colonial notions of other societies as savages.(Eaton, n.d) Through his films he conveyed the message that was given by Malinowski long back. He asked people to understand each society in their own unique context and to respect their values and institutions. Rouchs films are true representation of his thoughts on humanity and global understanding (Eaton,n.d). Introduction My earnest desire to write a Visual Anthropology essay on the works and contributions of Late Jean Rouch grew more profound after watching his film† Les Maites Fous† (The Mad Masters) .The film was released in the year 1955 and was a subject of great dispute among the various African intellectuals and French scholars. At the first glance I felt very uncomfortable with the scenes and visual presentations of the Film, but as I kept on watching it, true meanings of the film kept on unfolding in front of my eyes. One of the major reasons for choosing Jean Rouch and his film as the topic of my essay is because through my research I realized that most of the scholars and researchers have avoided critical evaluation and organized research   on Rouch’s films due to unavailability of the films and also because major number of films are in French. This has diminished the contribution of Jean Rouch and his films in the sector of Anthropological and Ethnographic films to an extent especially in the English Speaking Countries. The neglecting behavioral pattern towards Rouch’s films instigated me to find out more about Jean Rouch and his films. As Paul Stroller (1992) puts forward, that Jean Rouch is not given the right place in the history of anthropology and film and is often misunderstood and condemned by different scholars. In this essay I will talk about the various notions attached to Jean Rouch and his films, while critically evaluating his work â€Å"Les Maitres Fous†(The mad masters), 1955. Jean Rouch as a Film maker and an Ethnographer Jean Rouch saw cinema as an adventure and himself as an adventurer. As Claude Jutra(1960) comments â€Å" A chronicle of the Rouchian adventure is certainly an exciting prospect , but it is one that I approach with caution. One never admires without reservation. Any tribute carries within it an element of denunciation. No eulogy deserves to be trusted unless it is combined with certain degree of meanness†. (Jutra,1960:2). Jean Rouch dreamt of cinema as the only means of telling truth about people and societies. Stroller designated Rouch as the most prolific ethnographic filmmaker in the world whose ethnographic works are much unknown (Stroller,1992). Rouch clearly represented a unique combination of anthropology and art in his films. According to Brink, he was a filmmaker who did the most initiating job in pushing the boundaries of visual anthropology to an extent whilst still following the important norms and perceptions of visual anthropology of his time (Brink,n.d). Henley(2009) claimed that his films blurred the distinct lines between fiction and reality. As Jean Rouch himself, elaborately comments â€Å"For me as Filmmaker and ethnographer, there is practically no frontier between documentary film and fiction film. Cinema art of the double, represents a transition from the real world to the world of the imaginary, while ethnography, the study of other a peoples’ system of thought, involve s a permanent criss-crossing from one conceptual universe to another, a form of acrobatic gymnastics, in which losing your footing is the least of the risks†.(Rouch,1981:31) Rouch’s most remarkable films were Les Maitres Fous (The Mad Masters 1955),Moi Un Noir (1958),Jaguar (1967), andChronique d’un ete (1960).  Ã‚   Grimshaw and Eaton suggested that Jean Rouch’s films were much controversial in nature as well as did create a lot of debates among the scholars and intellectuals of younger generations. Surprisingly enough, among the anthropologists, there has been a substantial transformation in the approach towards Rouch’s films which has made it as one of the best examples of traditional ethnographic films. Stroller comments Jean Rouch as the Champion of the African Ethnography and the â€Å"Father of The Nigerian Cinema†(Stroller,1992).Brink states that Rouch gave primary focus to African people in his films, which made his cinema appealing and engaging to a limited number of audiences (Brink,n.d). According to Eaton, Rouch wanted his cinema not only to serve the purpose of anthropological education but also to use i t in much greater scale in order to understand the universal human interests and disciplines (Eaton,n.d). Henley remarked that Rouch believed in 100% participation in the events he filmed (Henley,2009). According to Edgar Morin, Rouch was a â€Å"Filmmaker Driver† who immerses himself into the life of his subjects and participants. Morin who was a well known French sociologist, worked with Jean Rouch in one of his most celebrated works â€Å"Chroinicle of a Summer† or â€Å"Chronique d’ un ete†in 1960. It was him who categorised Rouch’s films as â€Å"Cinema Verite†. Morin has explained this term as â€Å"there remain the most difficult, the most moving, the most secret (aspect of social life): wherever human feelings are at stake , wherever the individual is directly involved , wherever there are interpersonal relationships of authority, subordination , comradeship , love , hate in other words everything connected with emotion fabric of h uman existence . There lies the great terra incognita of the sociological or ethological cinema , of â€Å"Cinema Verite† . There lies its promised land (Morin, 1962:5) â€Å"Cinema Verite† is very much different from the direct cinema. Rouch viwed â€Å"Cinema Verite† as   â€Å"Cinema Provocation†. He explained that through this provocation a new form of cinema emerges which is more realistic than the real. The most interesting fact is that at first Rouch, himself was a great supporter of direct Cinema but with the time he got more indulged into the fictional films following the ethnological perception.   According to Michel Marie (1976), â€Å"Direct† in the technical sense of the term designates the synchronous recording of image and sound , the concept of direct cinema designates first of all a new technique of recording pro-filmic reality. This term replaced the more enigmatic cinema such as â€Å"Cinema Verite† at the beginning of the sixties, applies beyond a simple technique, to the whole new stream of practices which overthrew methods of film making that was previously standardized completely through the industrial model. Marie commented that â€Å"As opposed to industrial cinema, fictional and spectacular, in case of direct cinema the action to be filmed is void of an anterior status† (Marie,1976:35). â€Å"Cinema Verite† It is a kind of cinema where there is a good amount of conversation and interaction between filmmaker and his subject. In this form of cinema there may be also little provocations which can be noticed from the filmmakers side to his subjects (Feld,2003).   Rouch as a kid was very much influenced by the film â€Å"Nanook Of the North† (1922), which became an inspiration to the films that he made in later life. Brink states that Rouch inserted new forms and ways in the style of film making. With his work he blurred the difference between real and unreal, truth and fantasy and literally played with his lenses. It was interesting to note that his inquisitive nature and the use of new technology gave birth to a different genre of cinema which more or less became unique in its own ways.   As Mick Eaton (n.d) suggests that Rouch has tried to defend his film making practice in a very significant way, according to Rouch his presence during the film making is a very crucial factor not because people i.e. his subjects and participants will reveal themselves more openly and honestly out of trust and faith but more because they will lie and feel uncomfortable. It is the manifestation of this side of participants which will be regarded as more profound disclosure than anything â€Å"candid camera† or â€Å"living cinema† ever could disclose. Eaton followed the words of Jean Rouch believed that camera uniting with presence of its director acts as a catalyst to the whole process of film making.  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚  Ã‚   Thompson comments that Surre alism played a very important role in the development of Rouch’s films (Thompson,n.d). Surrealism as a cultural movement influenced the mind of Jean Rouch to a great extent. It was a philosophical movement which according to Henley became very popular in Paris during the world war. Jean Rouch claimed in 1967 that for him making a film or cinema is like a surrealist painting. Rouch was a bridge builder in true sense. He bridged the gap between film and anthropology along with an introduction to new techniques and style of film making. Brink comments that â€Å"The filmmaker who was educated as civil engineer became a true bridge builder between film and anthropology, film and art, fiction and documentary , Europe and Africa†(Brink,n.d:2) Grimshaw states that Rouch’s Anthropological films are mostly based on the sense of intuition and are idiosyncratic in approach. His films are a visual treat for the students of visual anthropology. Grimshaw strikes a comparison between the anthropological cinemas of Jean Rouch and Mac Dougall where she describes Mac dougall cinema as detailed and practical while Rouch’s anthropological films to be more idealistic and imaginative. According to Grimshaw , rouch was a visionary   who used his films as a way to interpret the complexities and connections between the world and humanity. His ethnographic films were very much influenced by the political situation at that time. Rouch was very much inspired by the different freedom movements by the colonial people during that time which brought huge political, economic and social transitions around the world saw the colonies as oppressive masters. According to Grimshaw, Rouch was not only supportive of these movements but also gave different humanitarian expressions to it (Grimshaw,2001). Rouch’s ethnographic sensibilities as a film maker and anthropologist was very much shaped by his individualistic nature, background and political circumstances in France during that time (Grimshaw,2001). Rouch as an Anthropologist was very much trained in the path Maussian ideologies a very well know French anthropologist during 1920s till 1940s. Henley comments that Rouch himself never met Mauss but he came to know about his contributions in Anthropology through his supervisor Marcel Griaule who was a student of Mauss himself. Maussian influence is very much evident in earlier works of Jean Rouch, especially in the films he and Rosefelder produced together during their expeditions. Inspired greatly by Mauss’s idea to use camera gave birth to Rouch’s lifelong interest in â€Å"salvage ethnography†(Henley,2009). The most significant and appealing part of Rouch’s journey is that though other leading ethnographic film makers such as Robert Gardner and Judith Mac Dougall (2006) have made films based on different continent. However, Rouch’s focus has always been on Paris and Africa, with more concentration on Niger.   This approach of his was mostly influenced by Griaule (1938) and Dieterlen (1987) who believed that it takes twenty to thirty years to understand and have a deep knowledge about a particular society. This view is significantly supported by Jutra’s comment â€Å"Rouch, the ethnologist! Rouch, the explorer! Rouch, the traveler ! mere appearances. Rouch has two very circumscribed habitats : Paris and Niger†¦ he has no desire to cross these boundaries that he has so carefully traced†¦. Rouch is sedentary, a stay – at – home.†(Jutra,1961:116) From all the works of Jean Rouch I will consider Les Maitres Fous as the starting point of his greater expression for universal humanity. This film is notable in ways that it establishes and evokes the contours of Rouch’s humanistic vision (Grimshaw,2001) â€Å"Les Maitres Fous† (The Mad Masters) This film was first screened in Paris in the year 1955. According to Grimshaw the small number of audience of African intellectuals and French anthropologist present in the premiere of the film in Musee De l’ Homme was largely antagonistic in its response to Rouch’s works. Marcel Griaule called for the fim to be destroyed; Africans present during the screening of the film rejected it as bigoted and insulting. Shortly afterwards British government banned this movie to be shown in the colonial territories of West Africa (Grimshaw, 2001). Today† Les Maitres Fous† is widely accepted and considered as a classic of contemporary cinema. After watching the film consecutively for the third time I realized that the film’s power to move and perturb the audience has not reduced at all with the passage of time. Grimshaw comments that â€Å"Les Maitres Fous† inaugurates the distinguished series of films, which Rouch made between 1954 and 1960. Some of the fi lms which were made during this time was Jaguar, â€Å"Moi Un Noir†and â€Å"Chronique d’un Ete†, the most important fact about all of these films was that all f them were based in different themes. It is one of the most significant films in the growth of Rouch’s works as an ethnographer (Eaton n.d). Feld (2003) illustrates that Les maitres Fous was the earliest departure from purely evocative cinemas into a more unreal approach to event structures. He claims that having experiencing a ceremony several times, Rouch thought of breaching the vital aspects and approaches them as dramatic narratives. According to Eaton Rouch was asked to make this film after him and his wife Jane had given lecture at the British council in Accra. Rouch was approached by some of the Hauka priests and initiates present in that lecture who approached him to film their Hauka Annual Ceremony. The priests wanted to record the event not only to preserve it as a film and memory but also so that it could be used I the ritual itself (Eaton,n.d) . Grimshaw reflects that the film basically documents the course of possession ceremony held during one Sunday by members of the Hauka sect working as migrant labourers in Accra. During 1920s there was a form of strong resistance against the French colonial rule by this Hauka cult who had moved from their villages to work in Kumasi and Accra , the commercial areas of British dominated Gold Coast (Grimshaw, 2001)  Ã‚  Ã‚  Ã‚   . According to Feld the Haukas were the most famous possession cult among the Songhay , that reached its full expression in Ghana, brought by the migrants from Niger. Mick Eaton explained that these ceremonies of Hauka sect started in the late 20s in the upper Niger region. Discriminated by the French colonial administration and adjudicated by rigid Islamic supporters many of the members of the Hauka sect moved to Ghana in the thirtees working as a migrant labourer throughout the gold coast region. Haukas defined themselves as the â€Å"new gods† the spir its of power and the winds (Eaton n.d) .The film vividly views the members of the Hauka cult working at menial task in the city during the week then in the possession trance during the weekend and again back to work in the week. The film starts with the scene of Bustling city of Accra where different people from across West Africa jostle up with one another sharing the â€Å"Great African adventure of work and life†. From the start the film appears to be imparting a sense of complexities, movements and characteristics of urban life.   Then there is sudden shift of events and audience find themselves plunged into a dark deep forest where Hauka people travelled by buses and trucks. According to   Rouch’s commentary   this was the place where the Hauka annual ceremony takes place and where the high priest of Hauka known as Mountebya lives.   The film shows the preliminary stages of confessions and appointment of new members before arriving into a dramatic climax w here members become possessed, adopting the identities and conventional behavior of British governor and his crew. The ceremony reaches its peak with the killing and eating of a dog, slowly Dusk approaches and Hauka people emerges from their trance and get prepared to return to Accra. The film ends with streams of flashbacks. Rouch is shown to return the following day to the same Hauka members who were possessed in the ceremony, calmy and efficiently working in their respective jobs (Grimshaw,2001). The possessed and frothing of the mouth, dribbling saliva flecked with the blood of the butchered dog, the violent and uncontrollable body movements and disturbing style of eating the dog meat are the most troubling factors of the film which shocks the senses of human mind and leaves a lifelong impression in ones’ mind. These shocking scenes of the film justify the fact behind the film being rejected by most of its audience as a whole. Indeed as James Clifford notes that this film stands as Rouchs’s manifestation of Griaule’s recognition that ethnographic observation involves conflict. According to Grimshaw â€Å"Les Maitres Fous†, reveals the sharp contrast between the conception of a shared or participatory anthropological cinema persued by Mac Dougall and the one practiced by Rouch†(Grimshaw,2001:101) â€Å"Les Maitres Fous† is distinguished for its intricate style camera work which Rouch adopted during his earlier life after the destruction of his tripod. Grimshaw (2001) also talks about Rouch’s narration in the Film as improvised or unrehearsed. It is his commentary that makes the film comprehensible and incomprehensible at the same time. Feld commented that Rouch made the film â€Å" Using a montage to create contexting boundaries and making the most of the technical limitation of twenty five second shots (he was still using a 16mm camera),Rouch was able to make a short film with more explicative depth and synthesis than his previous ethnographic studies†.(Feld,2003:5) As   Feld (2003) and Eaton (n.d) both expressed   that the film was shot on a hand cranked 16mm Bell and Howell Camera which allowed for 25 second shots, but it was also edited in the camera as much as possible and the ultimate shooting ratio turned out to be 8 – 10. The sound was recorded by Lam Ibrahim Dia and Damoure Zika, one of the first Africans whom Rouch knew quite well from his first trip to during the war, using a scubitophone which is a portable though heavy tape recorder with a clockwise motor that had to be wound up between takes.   Henley   added that the film is of 28 mins, edited by Suzzane Brown and produced by Les Films de la Pleiade and was awarded Grand Prix in the ethnographic category of the Venice Film festival in 1955 (Henley,2009). Eaton significantly estimated that Rouch’s use of voice over in â€Å"Les Maitres Fous† does not preserve the primacy of the images rather sets up completely different relation with sound and image. Rou ch not only translates the ceremony held by Huaka sect but more or less acts it out which in a way kills the authenticity of an ethnographic documentary (Eaton,n.d). Cooper (2006) suggests that though Rouch has tried to gain ethnographic knowledge through his ethnographic films as claimed by Jay Ruby (2000) but there is still a lot of space in acknowledging the fact which cannot be known or understood through the lenses. With no formal training in film making and direction, Rouch suprsing arose as a rule breaker in the Film world. Les Maitres Fous was one of the biggest examples of such rule break. According to Cooper† Les Maitres Fous† as a film can be represented in two ways at first considering the fact it is filmed by a western filmmaker it can be interpreted as a show of uncivilized attitude by the African Hauka people and secondly it can also be deciphered as African representation of the oppressive colonial western culture and its position in their society (Cooper ,2006). According to Homi.K .Bhabha the film is a clear representation of mockery to the oppressive colonial rule and their power show while showing the deep sighted pains of oppressed Africans (Bhabha,1994). The voice over used by rouch in the film is quite assertive and does influence the thinking process of the audiences a lot. Conclusion Rouchian cinema indeed consists of the most exciting form of ethnography which can be traced in all of his movies. My personal experience of Rouch’s film is a mixture of both the senses of agitation and fascination simultaneously. His film is indeed an experience of life time which stays in your memory till the last days of your life and which evokes the true spirit of humanity irrespective of class race and ethnicity. Through this Essay I have tried to discuss the various aspects of Jean Rouch’s cinema significantly concentrating my attention on â€Å"â€Å"Les Maitres Fous†. This movie disturbed my senses and my thinking about the human acceptance of culture. Through this film I realized that each society is different and is composed of different social dynamics. To me this film is a clear representation of the influence of oppressive western civilizations on the Africans and there after effects on African society. Though it should be also noted that this film has a element of fiction into it which sometimes covers the facts and presents a new form of truth which cannot be trusted sometimes. Cinema is the combination of rational, irrational, fiction and fact. Rouch expresses these elements more profoundly through his movies. Bibliography Henley, P .2009. The Adventure of the Real: Jean Rouch and the craft of Ethnographic Cinema. London and Chicago: The University of Chicago Press Ltd. Grimshaw, A . 2001. The Ethnographer’s Eye : Ways of seeing In Anthropology .New York , Cambridge and Melbourne: Cambridge University Press. Stroller, P.1992 .The cinematic Griot:the ethnography of Jean Rouch .London and Chicago: The University Of Chicago Press. Cooper , S .2006. Selfless Cinema?:Ethics and French Documentary. London: Legenda. Bhabha, H.K.1994. Bhabha: The Location Of Culture. New York: Routledge. Ruby ,J. 2000.Picturing Culture: explorations of film anthropology. Chicago and London:The University Of Chicago Press. Griaule , M. 1938. Masques Dogons. California:Institut dethnologie . MacDougall , D.2006. The corporeal image: film, ethnography, and the senses. Oxfordshire and New Jersey: Princeton University Press. Eaton, M.,n.d. Chronichle. In: Eaton. M,ed. 1979. Anthropology Reality Cinema: The films of Jean Rouch. Colchester and London:British Film Institute. p 1-34 Michel, M.,n.d. Direct. In: Eaton. M,ed. 1979. Anthropology Reality Cinema: The films of Jean Rouch. Colchester and London:British Film Institute. p 35-39 Eaton, M.,n.d. The production of Cinematic Reality. In: Eaton. M,ed. 1979. Anthropology Reality Cinema: The films of Jean Rouch. Colchester and London:British Film Institute.p 40-53 Rouch,J.,n.d. The Camera and Man. In: Eaton. M,ed. 1979. Anthropology Reality Cinema: The films of Jean Rouch. Colchester and London:British Film Institute.p54- 63 Brink.J.T.,n.d. Introduction. In.Joram .T.B,ed. 2007.Building Bridges: The cinema of Jean Rouch.London and New York :Wallflower Press.p 1 Nijiland,D.,n.d. Jean Rouch: A builder of bridges. In.Joram .T.B,ed. 2007.Building Bridges: The cinema of Jean Rouch.London and New York :Wallflower Press.p21 Grimshaw.A.,n.d.Adventure on the Road:Some reflections on Rouch and His Italian Contemporariries. In.Joram .T.B,ed. 2007.Building Bridges: The cinema of Jean Rouch.London and New York :Wallflower Press. Thompson.C.,n.d. Chance and Adventure in the Cinema and Ethnography of Jean Rouch. In.Joram .T.B,ed. 2007.Building Bridges: The cinema of Jean Rouch.London and New York :Wallflower Press Feld.S.,2003. Cinà ©-ethnography Jean Rouch.Minneapolis:University Of Minnesota Press

David Cole Interviews Dr. Franciscek Piper Essay - 1

David Cole Interviews Dr. Franciscek Piper - Essay Example But according to interview, Cole has disapproved the theory of concentration camps and gas chambers as gas used was just a disinfectant. Piper was an eye witness to the holocaust and believes that the figure projected by Russian was much higher, only about 1.1 million Jews had died as against the Russian propaganda of 4.4 million Jews who had been exterminated! The bullets marks were that of Air Force raids when it was turned into airbase for Germans while fighting against Allied forces. Cole believes that Auschwitz’ gas chambers and concentration camps were wild imaginations of few and Jews had probably died due to hard working conditions. But 1.1 million is not a small number and if Piper has quoted this number, then mass murders did happen. Whether they were killed or not in gas chambers or were tortured to death in concentration camps is immaterial. I fail to understand the purpose of Cole’s interview with Piper because even though, he has found no evidence that prove that people could have been killed in the gas chamber that was shown to Cole, the interview, nevertheless clearly reveals that such heinous crime was committed in Auschwitz by Germans in WWII. (words:

3 dyadic interview and appellations of concepts Essay - 2

3 dyadic interview and appellations of concepts - Essay Example However, I realized that they are much conserved and the information that they gave was not complete. Despite this, when giving their answers they were cool, calm, and collected. My focus was to know X’s interpersonal relations. This relates to the way they are able to interact with others as well as the extent to which they express their feelings as well as personal reaction to situations. When communicating with others X is very brief and precise, not that he is cautious of his words but at some point, he seems to lack the words. His expressions are not fully complete. However, he demonstrates clarity in the way he communicates. I talked with him concerning his ability to communicate with others. In this, he was very categorical that he finds it difficult to initiate conversations with strangers. Not only is this his predicament, but also maintaining a conversation with a stranger is a daunting task to him. Despite this, when communicating with a person he is conversant with , he pointed out that he is very free and can communicate for long time without any breakdown in communication. I yearned to know more about his interaction with family members and friends. He pointed out to me that have been with a single parent, who is his mother. Therefore, he points out that there are some things that he is not comfortable sharing with the mother. Being the only child, he has no option rather than learn from friends. His scope of friends is relatively small but his friends are very close to him. He demonstrates a deep sense of affection and does not take relationships lightly. He tells me that he has a girlfriend whom they spend a lot of time together. They have been together for the last three years. He tells me that his mother was initially opposed to the idea but she came to appreciate. As the conversation went on, he was easy going and could explain his answers in much more details compared to the

Analysis Cases In Arbitration Research Paper Example | Topics and Well Written Essays - 3250 words

Analysis Cases In Arbitration - Research Paper Example The U.S. Court of Appeals had declined to reassess the decision made by the en banc on February 1, 2012, in the case of Italian Colors Restaurant v. AET Related Services Co. Owing to this reason, the Supreme Court of the US is quite likely to be requested to revisit the issue related to class action waivers in the arbitration agreement by the US Court of Appeals. The dispute arose because of the fact that the Second Circuit denied offering an en banc rehearing. The chief judge, as well as four other judges, asserted that the case must be reheard which led to splits among the judges grounded on the protest that the Ninth Circuit was not being followed. In Italian Colors, it can be observed that the Second Circuit panel did not impose the class action waiver in relation to AE arbitration agreement considering that it would prevent the claimant from impeaching its federal antitrust claims. In the complaint, the Plaintiff claimed that the merchant contract disobeyed the SA. The merchant contract comprised of the arbitration provision that needed all claims that arose from agreement to be decided by means of arbitration. The SC on remand will face the concern of whether class-action waivers in case of arbitration agreements are unconscionable just because they prevent disputes from being resolved by ways of class proceedings. In an order list that was published within a week after the decision, it was found that the Supreme Court provided a writ of certiorari and thus evacuated and remanded the case of AE v. ICR.merican Express v. Italian Colors Restaurant. The court further ordered the Second Circuit to reassess its decision in relation to the unenforceability of class-action waiver in light of the Stolt-Nielsen opinion (Consumer Financial Service Group, â€Å"Second Circuit Splits with Ninth Circuit on Enforcement of Class Action

Determination of Sodium Thiopental Using Gold Nanoparticles

Determination of Sodium Thiopental Using Gold Nanoparticles Development of a new colorimetric method for the determination of sodium thiopental using gold nanoparticles Sodium thiopental (sodium pentothal) is in a group of drugs called barbiturates.this barbiturate commonly used anesthetic induction agents in man and animals because recovery is rapid and it has the advantage of having very little or no side effects[1].It is used for intensive-care patients with head injuries to control convulsions and reduce raised intracranial pressure[2]. As a resultmonitoring of theserum concentrations is important in this patient population. Several analytical procedures have been reported for the quantitative determination of thiopental. Among these high-performance liquid chromatography (HPLC) are more popular. HPLC assays are not completely reliable, and do not have the short process-time required in most of the above-mentioned indications[3, 4]. other methods are available for determining thiopental including stripping voltammetry[5],membrane sensors[6],capacitive chemical sensor [7],gas chromatography (GC)[8],spectrophotometric and spectrophotofluorometric[9, 10]. Donald et al[11]reported that, after the usual 4.8 mg/kg induction doses, thiopental concentration in serum as a function of time varies between 10 mg/L to 25 mg/L during 50h.As stated before most of these currently used methods for sodium thiopental detection usually need expensive and complicated instruments and are time-consuming, making on-site and real-time thiopental detection difficult. Therefore, it is important to develop a simple reliable and hig hly sensitive method for on-site and real-time detection of sodium thiopental. Recently, gold nanoparticles (NPs) explored for metallic NP-based colorimetric detection have attracted considerable attention due to biocompatibility, stability, and high extinction coefficients[12]. gold nanoparticles present size-dependent optical properties owing to the surface plasma resonance(SPR)[12]. The color of the colloidal Au NPs can be readily and precisely changed via aggregation of Au NPs.Au NPs were widely applied in colorimetric detection of several analytes such as protein, DNA, metal ions and small molecules[ ]. In this study, we used gold nanoparticles as a colorimetric probe for sensitive and selective detection of sodium thiopental. The gold nanoparticles were prepared using the classical citrate method [12].thiopental on the surface of AuNPs displaced the stabilizing citrate ions because thiol group of sodium thiopental tends to readily adsorb onto the surface of colloidal gold via chemisorptions-type interactions. The thiopental capped Au NPs were stable at basic and neutral conditions .Puntes et al [13] have studied the stability of cationic gold nanoparticle bioconjugates as a function of pH and the presence of citrate in solution. The pH of an aqueous solution of thiopental-Au NPs was varied by direct addition of citrate buffer. the thiopental-Au NPs can be aggregated by adding certain amounts of citrate buffer due to the electrostatic attraction between amino group contained in thiopental molecular and citrate ion on the surface of Au NPs, the amino group of the thiopental would be positively charged at the given pH value and they would therefore interact electrostatically with the negative charges of the citrate molecules. Thus forcing the aggregation of the conjugated Au NPs and subsequently resulting in the color change from wine red to purple or blue color.So that we detected it by UV–Vis spectrophotometer and paptode techniques and contrast both methods.First time at 2004 paptode was developed in Dr. Abbaspour group for speciation of iron(II) and iron(III) and the full range pH monitoring [14]. Then it was used for the determination of dopamine [15], hydrazine [16]. In paptode, conventional à ¯Ã‚ ¬Ã¢â‚¬Å¡atbed -scanner (as a nondestructive detector) was used to acquire the analytical parameters for quantitative determination of analyte that occurs via colorimetric reaction. The estimated re à ¯Ã‚ ¬Ã¢â‚¬Å¡ection density, as an analytical parameter, is obtained from an area of the sensing zone of spots using the average Red (R), Green (G) and Blue (B) channel. Degrees of the color of the spots are found to be proportional to the concentration of the testedanalyte. Experimental section: Reagents: HAuCl4.3H2O, trisodium citrate and citric acid were purchased from Sigma. Thiopental was obtained from Biochemie (Kundl, Austria) and zinc sulfate purchased from Fluka All solutions were prepared with ultrapure water Apparatus and software: The colorimetric study of NPs were performed by means of a Shimadzu 1601PC UV–Vis spectrophotometer (Kyoto, Japan)from 300 to 700 nm. Also a Canon scanner were used to record the color changes in paptode technique. The paptode Cells were built by creation of the holes (i.d 1.5 cm) in the sheet of plexiglas (thickness 0.9 cm). We used by photoshop Cs6 software to convert the recorded pictures of color of cells to RGB (Red, Green and Blue) and L*a*b data. The morphology and size of the nanoparticles were characterized by a transmission electron microscope (TEM model CM10; Philips). The X-Ray diffraction (XRD) patterns were obtained by using a D8 ADVANCE type (BRUKER-Germany) with Cu-KÃŽ ± radiation (ÃŽ »= 0.1542 nm). Powder XRD patterns were taken in 0.02 ° steps at 1 s per step. All the experiments were carried out at room temperature(25  ± 2 C) Synthesis of citrate-stabilized Au nanocrystals: Nanoparticles of noble metal were prepared by classical citrate method[12].the10ml of 0.014M of trisodium citrate dehydrate solution was added quickly to the 100ml of boiling solution of 0.5mM of HAuCl4.3H2O under magnetic stirring. The stirring was continued until a dark red color was observed (around 20 min) and the maximum absorbance of AuNPs solution was centered at 520 nm Sample preparation: Fresh human blood samples (2.0 mL) were obtained from volunteers of the local hospital. After letting sample stand for 60 min at room temperature we centrifuged at 4000 rpm for 10 min. The supernatant was used as the source of the serum. We used zinc sulfate method as a deproteinization technique[]: we vortex-mix for 10s of the 10ml of serum sample and 150mg zinc sulfate, then we centrifuged the mixture at 3000 rpm for 20 min. The supernatant, which excluded protein, was used for further analysis. Procedures for the detection of sodium thiopental: In a typical detection of sodium thiopental, different amounts of thiopental solution were added to the above XmlAu NPs solutions at room temperature. we proceeded to study the behavior of the conjugated system by modifying the pH . To investigate the effect of pH of the buffer solutions on thiopental detection, 0.5 mL of 0.1 M buffer solution (citric buffer solution in the pH range of 3.0–6.0 ) was added in mixture of thiopental and Au NPs solution. The obvious color change was observed with the naked eye and the absorbance spectra and scanning images of the solution were recorded 1 min after the addition of citrate buffer. In spectroscopy technique ,The concentration of sodium thiopental was quantified by the absorption ratio (A670/A520). Results and discussion Citrate was chosen as the stabilizer for AuNPs because it is negatively charged, and can act as a stabilizingagent to disperse AuNPs in aqueous solutions. The Au NPs after synthesis showed a surface plasmon resonance (SPR) band at 405 nm (Fig. 1a). the addition of sodium thiopental doesn’t led to a color change of Au NPsin ultrapure water, although the thiol group of sodium thiopental tends to readily adsorb onto the surface of Au NPs.The pH of AuNPs solution in present of sodium thiopental is 10.2 and Puntes et al[13]reportedthat the presence of charged molecules insolution may induce NPs aggregation by bridging particlestogether. It was observed that multiple electrostatic interactions between the conjugates mediated by cross-linking species led to an effective strong bond and consequently to irreversible aggregation and precipitation. So that at the given pH value , charge of thiopental can be change and thenthe color of the colloidal thiopental-Au NPs can be changed to blu e (broad band above 600 nm).*Scrutiny of pH/Concentrate diagrams of citrate and thiopental shows that at the pH of between 5 to 7 , charge of citrate and thiopental can benegative and neutralfig S1. But when sodium thiopental add to AuNPs solution, the S- group in the sodium thiopental provides a strong affinity for gold. So that orbital of thiol group of thiopentalinvolved for Au NPs surface and when pH change from 10.2 to 6 , the amino group of the thiopental would be accepted H + and get positive charge. In present of excesscitrate at the pH of 6 , thiopental-AuNPscan be aggregated via electrostatic attraction between the citrate ions and the thiopental. So that in this study we used citrate buffer solutionfor control of pH( in the pH range of 3.0–6.0) and source of citrate (as a bridging factor). The aggregation mechanism of Au NPs is illustrated in Fig. 1. Optimization pH and time we proceeded to study the behavior of the conjugated system by modifying the pH( 7.1-5.4). The pH of an aqueous solution of0.00001M thiopental capped AuNPs was varied by direct addition of 0.05Mcitrate buffer to the solution andThe UV-Vis spectrum wasmonitored and the extinction ratio of absorbance at 600 nm to 420 nm (A600/A410) is plotted against the pH inFig. 3A. The thiopental-capped Au NPs were stable at basic and neutral conditions.When the pH of the solution was below the 6.4 , Au NPs agglomerated.the aggregation was solely due to the bridging citrate between the amine functionality.Onthe basis of this optimization experiment, the pH was set to 6.2 to achieve a best aggregationFig. 3A.When the pH was decreased immediately from 5.4 after the addition of the citrate buffer scatteringwasobserved.Fig. 3A illustrates theabsorption spectra of AuNPs at different pH value. At the concentration of sodium thiopental as 0.00001M, the extinction ratio ofA650/A520 at room temperatureexhibited a rapid increaseduring the first 1.5min,then increased gradually from 1 min to 18 min and then remained constantFig 3B. Thus, the detection time was chosen as 20 min. We choseto use the absorbance ratio at 500 and 600 wavelengths to quantify thecolor of the system,thecolor change at various sodium thiopental concentrations were monitored byUV/Vis spectroscopyfig4A.Quantitative analysis was performed by monitoringthe absorbanceat 1minute after the addition of citrate buffer Fig4B .The linear range, detection limit and reproducibilityof the method were evaluated under the optimumconditions.Thecalibration curve for sodium thiopental was linear in two ranges of( †¦Ã¢â‚¬ ¦. To †¦Ã¢â‚¬ ¦ and†¦Ã¢â‚¬ ¦ to †¦Ã¢â‚¬ ¦) with correlation coefficients 0.9981 and0.9979, respectively. The Experimental detection limit has been obtained as 2 µM. The relative standard deviation(R.S.D.) for1.0Ãâ€"10−8M thiopental measurementwas2.7% (n=11)Fig4A .when thiopental concentrationincreased above 0.0005M, scattering was observed fig3B because thiopental polymerized white citrate molecule. So that we tried paptode techniques to resolve thisproblem FigS1. Although the higher concentrations of sodium thiopental was determined by paptode, but the limit of detection was rather high (LOD 10  µM) in comparison to the spectrophotometric method. The detailed procedure for sodium thiopental determination by the paptode method is explained in supporting information. To test the selectivity of the above method for sodium thiopental, we testing the response of the assay to some potential interference species and structurally similar to the sodium thiopental such as†¦Ã¢â‚¬ ¦Ã¢â‚¬ ¦Ã¢â‚¬ ¦.in optimum condition and different concentration .the results areshown in bar diagramFigure 8 .red barsexhibit Color changes of the solution in thepresence of various interference species at concentrations of 10mMand bluebars exhibit Color changes in presence ofinterference species at real concentration in serum ( 1M cysteine, 2M†¦Ã¢â‚¬ ¦Ã¢â‚¬ ¦), The maximum absorption wavelength of AuNPs did notchange in the presence of the tested species, Except for cysteineat concentrations of 10mM. Therefore, AuNPs had good selectivity for sodium thiopental detection in optimum condition in the serum. Colorimetric detection of sodium thiopental in serum: To validate the reliability of the proposed method for sodium thiopental detection in real samples, The unknown amounts of thiopental were added to thethree different human serum samples before samplespre-treatment .Detecting of sodium thiopental in a serum is not easy because of the serum constituents.the color of the Au NPs was not stable by the addition of the blank serum. So that it mustdiluted ten times. As regardsthe calibration curve for detection thiopental by this methodand dilution of serum and thiopental concentration in serum as a function of time varies after the usual 4.8 mg/kg induction doses [] , we can detect sodium thiopental in human serumbefore 3 hour.samples were determined by both the AuNP-based method reported herein and the standard addition method. Satisfactory results and recoveries as shown in Table 2. The satisfactory results obtained indicate that proposed sensors can be applied to real sample assays. 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