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Evaluation of a new coprocessed compound based on lactose and maize starch for tablet formulation.

The development of new direct compression excipients should include a comprehensive and rapid determination of deformation properties. The aim of this study was to characterize StarLac, a new coprocessed compound for direct compression based on lactose and maize starch. For this purpose, the effects of the base materials (maize starch and spray-dried lactose) were considered and the influence of the spray-drying process was investigated. This was performed by comparing the physical mixture of starch and spray-dried lactose at the same ratio as for StarLac. For analysis of the deformation behavior, the 3-D model and the Walker equation were applied; for verification, the Heckel equation and the pressure time function (a modified Weibull equation) were used. The advantages of StarLac are its good flowability depending on the spray-drying process, an acceptable crushing force due to its lactose content, its rapid disintegration depending on starch, and a brilliant fast release of an active ingredient, such as theophylline monohydrate. The volume-pressure deformation properties of StarLac were dependent on the lactose properties. Only at high maximum relative density (rho(rel, max)) did the influence of starch cause a change in these properties. A network-like structure can be observed using scanning electron microscopy pictures. Overall, StarLac deformed plastically with a low portion of elasticity. The physical mixture exhibited a more elastic behavior than StarLac. However, the part of the powder that was irreversibly compressed was much lower than was observed for the single substances. This behavior is caused by an interaction between the components, which in StarLac is prevented by spray drying.

Compressive Strength↗

Effects of physical properties for starch acetate powders on tableting.

The aim of the study was to investigate particle and powder properties of various starch acetate powders, to study the effect of these properties on direct compression characteristics, and to evaluate the modification opportunity of physical properties for starch acetate powders by using various drying methods. At the end of the production phase of starch acetate, the slurry of starch acetate was dried using various techniques. Particle, powder, and tableting properties of end products were investigated. Particle size, circularity, surface texture, water content and specific surface area varied according to the particular drying method of choice. However, all powders were freely flowing. Bulk and tapped densities of powders varied in the range of 0.29 to 0.44 g/cm3 and 0.39 to 0.56 g/cm3, respectively. Compaction characteristics revealed that all powders were easily deformed under compression, having yield pressure values of less than 66 MPa according to Heckel analysis. All powders possessed a significant interparticulate bond-forming capacity during compaction. The tensile strength values of tablets varied between 10 and 18 MPa. In conclusion, physical properties of starch acetate could be affected by various drying techniques. A large specific surface area and water content above 4% were favorable properties by direct compression, especially for small, irregular, and rough particles.

Excipients↗

Gelatin and hydroxyethyl starch, but not albumin, impair hemostasis after cardiac surgery.

We investigated the effect of postoperative administration of colloids on hemostasis in 45 patients after cardiac surgery. Patients were randomized to receive 15 mL kg(-1) of either 4% albumin, 4% succinylated gelatin, or 6% hydroxyethyl starch (molecular weight of 200 kDa/degree of substitution 0.5) as a short-term infusion. There was a comparable decrease in maximum clot firmness of thromboelastometry tracings in gelatin and hydroxyethyl starch groups immediately after completion of the infusion, whereas these values remained unchanged in the albumin group. The impairment in clot strength persisted up to 2 h, although the values partly recovered. Postoperative bleeding correlated inversely with the clot strength in pooled data of the artificial colloids. Fibrin formation (clot formation time, alpha-angle) and fibrinogen-dependent clot strength (maximum clot firmness and shear elastic modulus) were more disturbed in the hydroxyethyl starch group than in the gelatin group. We conclude that after cardiopulmonary bypass surgery, both gelatin and hydroxyethyl starch impair clot strength and fibrin buildup, which may predispose patients to increased blood loss. The greatest impairment in hemostasis was seen after hydroxyethyl starch administration, whereas albumin appeared to have the least effect on hemostatic variables.

Adult↗

Purification and characterization of a thermostable raw starch digesting amylase from a Streptomyces sp. isolated in a milling factory.

A raw starch utilizing microbe was isolated from mud in a milling factory. The 16S ribosomal DNA (rDNA) sequencing and morphological properties of the strain indicated that it belongs to the genus Streptomyces. A strongly raw starch digesting amylase was purified from the culture supernatant of the strain by chromatographic procedures. The specific activity of the enzyme was 11.7 U/mg, molecular mass 47 kDa, optimum pH 6.0, and optimum temperature 50 to 60 degrees C. The enzyme showed sufficient activity even at 70 degrees C. It was activated by calcium, cobaltous, and magnesium ions, and inhibited by copper, nickel, zinc, and ferrous ions. It formed maltose mainly from raw and gelatinized starch, and glycogen. No products were formed from glucose, maltose, maltotriose, pullulan, or cyclodextrins (CDs). The enzyme digested raw wheat, rice, and waxy rice starch rapidly, and raw corn, waxy corn, sweet potato, tapioca, and potato starch normally.

Amino Acid Sequence↗

Effect of bread containing resistant starch on postprandial blood glucose levels in humans.

We examined the inhibitory effect of a single ingestion of bread containing resistant starch (bread containing about 6 g of resistant starch derived from tapioca per 2 slices) (test food) on the postprandial increase in blood glucose in male and female adults with a fasting blood glucose level between 100 and 140 mg/dl. Bread not containing resistant starch (placebo) was used as the control. The study was conducted in 20 subjects (9 men and 11 women with a mean age of 50.5+/-7.5 years) using the crossover method, with a single ingestion of either bread containing resistant starch or the placebo. Blood glucose and insulin were measured before ingestion, and at 0.5, 1, 1.5, and 2 h after ingestion. The blood glucose level before ingestion was stratified into a borderline group (blood glucose level >/= 111 mg/dl) and a normal group (blood glucose level </= 110 mg/dl), with the upper limit of the normal range defined as 110 mg/dl. Postprandial increases in both blood glucose and blood insulin were significantly inhibited in subjects in the borderline group who took the test food in comparison with the placebo group (blood glucose: p<0.05 and p<0.01 at 1 and 1.5 h after ingestion respectively; insulin: p<0.05, p<0.01 and p<0.05 at 1, 1.5, and 2 h after ingestion respectively). These results indicate that bread containing resistant starch is useful for prevention of lifestyle-related diseases such as diabetes mellitus, and as a supplementary means of dietetic therapy.

Adult↗

Influence of ruminal protozoa on site and extent of starch digestion and ruminal fermentation.

Eight ruminally, duodenally, and ileally fistulated sheep were used to study the effects of protozoa on ruminal fermentation and starch digestion in the rumen and intestines. The experimental diet (75% grain DM basis) was based on a mixture of 67% high-moisture corn (HMC) and 33% dry-rolled grain sorghum (DRGS). Protozoa were reduced from 51,286 to 13,987 organisms/mL of ruminal fluid by feeding a diet (13 d) containing 9% tallow and monensin at 27 mg/kg (DM basis). Three sheep were defaunated by intraruminal infusions of lactic acid (20 to 30 g/d) and one sheep by washing the rumen and heating ruminal contents (60 degrees C for 20 min). No differences were observed in DM or starch intake, ruminal fluid pH, lactate concentration, or total tract starch digestibility. Molar proportion of acetate was reduced (48.6 vs 58.3%; P < .01) and propionate was increased (32.2 vs 23.4%; P < .03) by defaunation. Total VFA tended (P = .20) to be lower in faunated sheep (118.5 vs 139.2 mM). In situ rate of starch digestion of HMC (22.3 vs 14.7 %/h; P < .02) and DRGS (5.1 vs 3.3 %/h; P < .009) was increased by defaunation. Starch digestibility (percentage of intake) in the rumen was increased (P < .001) by defaunation (84.2 vs 93.7%) and was reduced (P < .002) in the small intestine (5.2 vs 13.6%). Defaunation increased (P < .03) amylolytic activity (.40 vs .17 units/mg of protein; P < .03) and osmotic pressure (321 vs 245 mOsm/kg; P < .006).(ABSTRACT TRUNCATED AT 250 WORDS)

Ammonia↗

In vitro fermentation of various fiber and starch sources by pig fecal inocula.

Freeze-dried ileal effluent (1% wt/vol) from cannulated pigs fed rice-based diets with the inclusion of either animal protein (CON), animal protein plus potato starch (PS), animal protein plus sugar beet pulp (SBP), or animal protein plus wheat bran (WB) was incubated anaerobically at pH 6.0 in fermenters containing 5% (wt/vol) fecal slurry comprising mineral salts medium and 50 g/L of fresh feces from pigs fed the same diets as the cannulated pigs. Samples were collected from the fermenters at 0, 2, 4, 12, 24, and 48 h during in vitro fermentation for measuring nonstarch polysaccharides (NSP), starch, and short-chain fatty acids (SCFA). Results showed that the major SCFA produced were acetate, propionate, and butyrate. The inclusion of soluble dietary fiber (diet SBP) caused the highest concentrations of acetate, propionate, butyrate, and total SCFA, whereas the increase in the production of propionate resulting from the addition of insoluble dietary fiber (diet WB) only occurred at the initial stages during 48 h in vitro fermentation. At all sampling occasions (except for 4 h), the levels of butyrate were increased (P < 0.01) by resistant starch compared with fiber sources, showing that a higher level of butyrate can be achieved through microbial fermentation by potato starch. Lowered (P < 0.05) butyrate concentrations were observed with diet WB during in vitro fermentation. With the inclusion of fiber sources, the energy originating from SCFA was similar to that from NSP disappearance, whereas the values were lower (P < 0.05) from NSP disappearance than for SCFA generated without fiber sources supplemented. We conclude that more substrate is available in ileal effluent with the addition of soluble dietary fiber, and an increased level of butyrate could be achieved through microbial fermentation by resistant starch.

Animals↗

Effect of starch infusion at the terminal ileum on nitrogen balance and apparent digestibilities of nitrogen and amino acids in pigs fed meat-and-bone and soybean meal diets.

Growing pigs were each surgically fitted with a single T-shaped cannula at the terminal ileum, and fed a cornstarch-based diet made isonitrogenous with meat-and-bone meal (MBM) or soybean meal (SBM). Starch or water was infused through the cannulas and the effects on N balance and apparent digestibilities (AD) of N and amino acids (AA) were studied. The infusion of starch, as contrasted to water, increased (P less than .01) excretion of fecal N and decreased (P less than .05) excretion of total urinary N, including urinary urea N, without affecting the amount of N retained. The increased excretion of fecal AA following starch infusion resulted in lowered (P less than .05) AD (percentage units) of indispensable AA, threonine (8.4), methionine (6.7), valine (6.6) and lysine (5.0); and dispensable AA, tyrosine (6.9) and aspartic acid (5.7). The same pattern of AD decreases was obtained for both protein sources, but in general, the magnitude of these decreases was greater for MBM than for SBM. Large differences (percentage units) were observed between AD of individual AA and that of N (arginine, +11.2 and isoleucine, -5.2, in MBM; arginine, +7.6 and methionine, -7.6, in SBM). Results showed that the route of N excretion and the AD of N and AA, as determined by the fecal analysis method, were influenced by the amount of starch entering the hindgut. It was concluded that AD of individual AA, and not N, should be considered when formulating pig diets; however, because of the dependence of AD values on starch entering the hindgut, AD values (fecal analysis method) may not be reliable measures of AA availabilities.

Amino Acids↗

Comparative study of the effects of equal amounts of fat, protein, and starch on plasma cholecystokinin in man.

The effect of ingestion of 50 g fat, 50 g protein, and 50 g starch on plasma cholecystokinin (CCK) concentrations was studied in eight healthy volunteers. Plasma CCK concentrations were measured by radioimmunoassay with Bolton-Hunter-labelled CCK 33, CCK 33 standard, and antibody T204. Antibody T204 was directed to the sulphated tyrosine region of CCK. Ingestion of fat and protein induced significant increases in plasma CCK, whereas ingestion of starch did not significantly influence plasma CCK levels. Peak increments in plasma CCK after fat (4.8 +/- 0.9 pmol/l) and protein (3.4 +/- 0.5 pmol/l) were significantly greater than that after starch (0.9 +/- 0.3 pmol/l). Similarly, the integrated plasma CCK secretion after fat (213 +/- 49 pmol/l X 120 min) and after protein (178 +/- 53 pmol/l X 120 min) was significantly greater than that found after ingestion of starch (9 +/- 23 pmol/l X 120 min). It is concluded that, in contrast to starch, fat and protein are potent stimuli for the release of CCK.

Adult↗

Assimilation of wheat starch in patients with chronic pancreatitis. Positive effect of enzyme replacement.

Pancreatic insufficiency due to chronic pancreatitis may lead to symptomatic malabsorption of both starch and fat. The absorption capacity of wheat starch has not been studied previously in patients with chronic pancreatitis, although this carbohydrate is a quantitatively important component of the Western diet. We studied the absorption of wheat starch and the effect of pancreatic enzyme substitution in seven patients with chronic pancreatitis and steathorrea. The malabsorption was determined from hydrogen breath tests with lactulose standards as reference. Without enzyme substitution, wheat starch (50 g) was absorbed to a lesser extent than in healthy controls (p less than 0.05). The mouth-to-cecum transit time was prolonged and correlated positively to the fat excretion before substitution with pancreatic enzymes (sigma = 1). The enzyme substitution increased the absorption of wheat starch to values seen in healthy controls (p less than 0.05) and reduced the mouth-to-cecum transit time by 19.8%.

Adult↗

Relationship between corn vitreousness and ruminal in situ starch degradability.

The objective of this experiment was to determine the relationship between corn kernel vitreousness and ruminal in situ starch degradation. Fourteen U.S. and five Brazilian corn hybrids cultivated in their respective countries were evaluated. The U.S. dent hybrids were harvested at one-half milk line, black layer, and 21 d after black layer stages of maturity. Brazilian flint hybrids were harvested only at the latest stage of maturity. Vitreousness was determined by manual dissection of the kernels. Ruminal in situ degradation of starch was determined in three lactating Holstein cows fitted with ruminal cannulae. Vitreousness of the five mature Brazilian hybrids averaged 73.1% (range of 64.2% to 80.0%), while vitreousness of the 14 mature U.S. hybrids averaged 48.2% (range of 34.9% to 62.3%). Within the 14 U.S. hybrids, average vitreousness increased from 42.8% to 48.2% as stage of maturity progressed from one-half milk line to 21 d after black layer. The correlation between kernel density and vitreousness was 0.87. The correlations between kernel vitreousness or density and ruminal starch availability were -0.93 and -0.87, respectively. With advancing maturity, kernel vitreousness and density increased while ruminal starch availability decreased. Kernel vitreousness and density may be useful parameters for which to select corn hybrids for high ruminal starch availability. Density may be a more practical measurement than vitreousness for screening large corn data sets.

Animal Feed↗

Varying protein and starch in the diet of dairy cows. I. Effects on ruminal fermentation and intestinal supply of nutrients.

The main objective of this experiment was to examine the effects of the percentage and source of crude protein (CP) and the amount of starch in the diet of dairy cows on ruminal fermentation, nutrient passage to the small intestine, and nutrient digestibility. For this purpose, 6 multiparous Holstein cows fistulated in the rumen and duodenum that averaged 73 d in milk were used in a 6 x 6 Latin square design with a 2 x 3 factorial arrangement of treatments. Two sources of CP [solvent-extracted soybean meal (SBM) and a mixture of SBM and a blend of animal-marine protein supplements plus ruminally protected Met (AMB)] and 3 levels of dietary protein (about 14, 16, and 18%) were combined into 6 treatments. On a dry matter (DM) basis, diets contained 25% corn silage, 20% alfalfa silage, 10% cottonseed, 26.7 to 37% corn grain, and 4 to 13.5% protein supplement. Intakes and digestibilities in the rumen and total tract of DM, organic matter, acid and neutral detergent fiber were unaffected by treatments. Increasing dietary CP from 14 to 18% decreased the intake and apparent ruminal and total tract digestion of starch, but increased the proportion of starch consumed by the cows that was apparently digested in the small intestine. At 14% CP, starch intake and total tract digestion were higher for the AMB diet than for the SBM diet, but the opposite occurred at 16% CP. Across CP sources, increasing CP in the diet from 14 to 18% increased the intakes of N and amino acids (AA), and ruminal outflows of nonammonia N, nonammonia nonmicrobial N, each individual AA except Met, total essential AA, and total AA. Across CP percentages, replacing a portion of SBM with AMB increased the intake of Met and Val and decreased the concentration of ammonia N in the rumen, but did not affect the intake of other essential AA or the intestinal supply of any essential AA and starch. The ruminal outflow of microbial N, the proportional contribution of Lys and Met to total AA delivered to the duodenum, and milk yield were unaffected by treatments. Data suggest that the intake of N by high-producing dairy cows that consume sufficient energy and other nutrients to meet their requirements can be decreased to about 600 to 650 g daily without compromising the supply of metabolizable protein if the source and amount of dietary CP and carbohydrate are properly matched.

Animal Nutritional Physiological Phenomena↗

Influence of dietary protein and starch on production and metabolic responses of dairy cows.

The effects of starch and CP degradability on ruminal digestion, milk yield, and milk composition were evaluated using a 4 x 4 Latin square design. Thirty-six Holstein cows (15 multiparous; 21 primiparous) and 4 ruminally cannulated cows were assigned randomly to four diets. The experiment was conducted during wk 4 through 16 postpartum. Diets contained 60% concentrate and 40% bromegrass hay. Treatments were barley plus canola meal and soybean meal; barley plus corn gluten meal, fish meal, and meat and bone meal; corn plus canola meal and soybean meal; and corn plus corn gluten meal, fish meal, and meat and bone meal. Intake of DM was not influenced by starch source, but CP intake was higher for diets based on corn than for those based on barley. Yields of milk, milk protein, and lactose were higher for diets based on fish meal than canola meal but were not altered by dietary starch source. Milk protein percentage and 4% FCM yield were not altered by dietary protein source. The 4% FCM and milk fat yields were higher for cows fed diets based on corn than for cows fed diets based on barley. Cows fed diets based on corn had significantly lower concentrations of most short- and medium-chain fatty acids and elevated concentrations of C18:0, C18:1, and C18:2 in milk. Protein source significantly influenced milk yield, and milk composition was altered by starch source. However, no interaction was observed between starch and protein sources.

Animal Feed↗

Effect of percentage of dietary forage neutral detergent fiber and source of starch on performance of lactating Jersey cows.

Five Jersey cows were used in a 5 x 5 Latin square design to determine the effects of decreasing dietary forage neutral detergent fiber (NDF) and different sources of dietary starch on performance and nutrient digestibilities. The control diet was balanced to consist of 21% forage NDF and 43% nonfiber carbohydrates. Four other diets were balanced to contain 35% nonfiber carbohydrates and either 16 or 11% forage NDF; diets were arranged in a 2 x 2 factorial design with either corn or corn and wheat as the sources of starch. Dry matter intake decreased linearly as forage NDF decreased; however, most of the decrease occurred when forage NDF was reduced from 16 to 11%. Milk production, yield of 4% fat-corrected milk, and percentages of milk fat and protein were similar among diets. Digestibility of NDF and acid detergent fiber increased as forage NDF decreased, but fiber digestibilities decreased with the addition of wheat to the diets. Starch digestibility was similar among diets. Source and amount of starch may be equally important or more important than the percentage of forage NDF for maintaining nutrient digestibilities of the total tract. Forage NDF in the diets of high producing cows can be reduced to 16% when sources and concentrations of starch are adequately balanced.

Animal Feed↗

Native corn starch versus either cellulose or glucose in the diet and the effects on apparent magnesium absorption in goats.

The objective of this study was to determine whether dietary glucose or starch would reduce the inhibitory effect of high K intake on Mg absorption in ruminants. In a 6 x 6 Latin square design, six goats were fed diets with or without added KHCO3 containing either cellulose, glucose, or native corn starch. The K concentrations of the diets were 7.8 or 34.0 g or K/kg of dry matter, and carbohydrates were incorporated so that their concentrations were identical on a glucose equivalent basis (331 g of glucose/kg of dry matter). The intake of extra K significantly reduced apparent Mg absorption from 29.8 to 22.1% on average. Glucose, instead of cellulose, in the diet did not affect Mg absorption. Replacement of dietary cellulose by corn starch enhanced the mean efficiency of Mg absorption from 21.8 to 30.9%. Starch versus glucose increased Mg absorption by 5.8 percentage units. No statistically significant interaction was observed between the type of carbohydrate and the amount of K in the diet with regard to Mg absorption. This study showed that the inhibitory effect of dietary KHCO3 on Mg absorption in goats was fully counteracted by the replacement of cellulose with starch in the diet. Possible changes in the pH of ruminal fluid might have mediated the dietary effects on Mg absorption.

Absorption↗

Some properties of starches of opaque-2, sugary-2 opaque-2, and waxy opaque-2 mutants of two broad-based synthetic cultivars of maize.

Starches of the opaque-2, sugary-2, opaque-2, and waxy opaque-2 endosperm mutants of two broad-based synthetic cultivars (Temp HA and Temp HB) of maize adapted to the more temperate areas of the world are, in general, similar to the respective starches of inbred maize background with regard to amylose percentage and the distribution of linear alpha-D-(1 leads to 4) linked unit-chains of amylopectin, starch-granule susceptibility to amylases, and properties examined by X-ray diffractometry and photopastegraphy. Changes in starch content and morphology of starch granules by scanning electron microscopy in dissected endosperms of the two broad-based synthetic maize cultivars following germination 0, 2, 4 and 6 days after planting were also investigated.

Amylases↗

Starch digestion in fowl.

Starch is usually the largest single nutrient in feed and provides the greatest proportion of metabolizable energy. Amylose and amylopectin comprise starch and are packed by plants in granular form. Granule stability is a function of the proportions of each polymer and the manner in which they are crystallized. Plant source determines granule size and stability. Grains generally have granules that are smaller and less stable than tuber or legume sources. Pancreatic alpha-amylase is the only enzyme elaborated by fowl that digests starch. Avian and mammalian sources are very similar, and inhibitors would not ordinarily be encountered in practice. The primary products of amylose digestion are maltose and maltotriose which further include alpha-limit dextrins when amylopectin is the substrate. Having starch in granule form reduces polymer access by the enzyme, and digestion difficulties occur in proportion to stability of structure. Moisture combined with heat destabilize granule structure and their use at one time or another in the manufacture of most feeds alleviates digestibility problems. Large amounts of the enzyme are present with the chick at hatch, and the pancreas is more than capable of synthesis commensurate with need. Gelatinization, enzyme adequacy, predominance of starch from grain, and lack of inhibitors account for the relative absence of practical problems involving this nutrient.

Amylopectin↗

Results of use of metformin and replacement of starch with saturated fat in diets of patients with type 2 diabetes.

OBJECTIVE: To improve glycemic control by substituting saturated fat for starch, to identify any adverse effect on lipids masked by the extensive use of metformin and lipid-lowering drugs, and to attempt to separate dietary effects from effects of multiple drugs. METHODS: We undertook a retrospective review of medical records of patients who completed 1 year of follow-up after dietary prescription. The study subjects included 151 patients in the diet group (whose dietary instructions included high saturated fat but starch avoidance) and 132 historical control subjects (who were allowed unlimited monounsaturated fat but had restriction of starch in their diets). RESULTS: Hemoglobin A1c (HbA1c) levels improved in both study groups (-1.4 +/- 0.2% [P<0.001]; 95% confidence interval [CI], -1.9 to -0.9). Use of metformin was associated with a decrease in HbA1c (-0.12 +/- 0.003%/mo [P<0.001]; 95% CI, -0.17 to -0.07). The diet group had an additional decrease of -0.7 +/- 0.2% (P<0.001; 95% CI, -1.1 to -0.3). Weight increase was associated with the use of insulin (+0.3 +/- 0.07 kg/mo [P<0.001]; 95% CI, 0.2 to 0.5), sulfonylurea (+0.18 +/- 0.06 kg/mo [P<0.01]; 95% CI, 0.05 to 0.30), and troglitazone (+0.7 +/- 0.2 kg/mo [P<0.005]; 95% CI, 0.3 to 1.2). Although not statistically significant, metformin therapy showed a trend for weight loss (-0.14 +/- 0.08 kg/mo; P = 0.07). An additional weight loss was noted in the diet group (-2.65 +/- 0.62 kg [P<0.001]; 95% CI, -3.87 to -1.44). Hydroxymethylglutaryl-coenzyme A reductase inhibitor use was associated with reduced total cholesterol level (-1.7 +/- 0.6 mg/dL per month [P<0.005]; 95% CI, -2.9 to -0.5). The diet group had an additional decrease of -13.0 +/- 4.5 mg/dL (P<0.001; 95% CI, -21.9 to -4.1). No significant effect of the diet on triglyceride, low-density lipoprotein, or high-density lipoprotein levels was detected. CONCLUSION: Addition of saturated fat and removal of starch from a high-monounsaturated fat and starch-restricted diet improved glycemic control and were associated with weight loss without detectable adverse effects on serum lipids.

Cholesterol↗