Cholesterol Extract Research Articles

Niosome gels encapsulate green mangosteen peel extract (Garcinia mangostana L.) as an anti-acne-inducing bacterial and anti-inflammatory activity

This research aims to develop a niosome gel from green mangosteen peel extract (Garcinia mangostana L.) by maceration with an ethanol solvent. The result of the yield percentage was 14.35 ± 0.90. The result of analyzing the phytochemicals by high-performance liquid chromatography (HPLC) was that tannin and xanthone were equal to 0.7483 ± 0.0825 mg per 100 mg of extract and 0.02964 ± 0.0088 mg per 100 mg of extract, respectively. The results of the determination of the effect include anti-inflammatory and anti-bacterial (Cutibacterium acnes) as anti-inflammatory with nitric oxide from LPS-induced macrophage cells up to a maximum equal to 29.10 ± 4.78% as a concentration at 1 mg mL–1 as acetonide can inhibit nitric oxide equal to 33.12 ± 3.62% as a concentration at 1 mg mL–1. and anti-bacterial (Cutibacterium acnes) by broth microdilution and drop plate methods, it was found that clindamycin has a minimum inhibitory concentration (MIC) and a minimum bactericidal concentration (MBC) of 0.08 μg mL–1 and 0.31 μg mL–1, respectively. The development of niosomes consisting of cholesterol, tween 60, and mangosteen peel extract at 1% w w–1 and 2% w w–1 in every formula showed good stability. The reduction of particle size in the formula by an ultrasonic bath at 30 minutes and measurement of particle size with a transmission electron microscope (TEM) were found to be equal to 1,265 and 123 – 219 nm, respectively. The polydispersity index (PDI) was in the range of 0.1 – 0.2, and the zeta potential value was in the range of –26.15 to –28.61 mV. The result of hydration and trans epidermal water loss (TEWL) was found to be that after 4 weeks of use, the formula containing the niosomes of mangosteen peel extract concentrated at 2% w w–1 maximizes skin moisture. It has a value of 346 ± 39.27 and has the least surface water loss. GRAPHICAL ABSTRACT HIGHLIGHTS The green mangosteen peel extract (Garcinia mangostana L.) can inhibiting nitric oxide production from LPS-induced macrophage cells up to 29.10 ± 4.78% at a concentration of 1 mg ml-1, the lowest inhibitory (MIC) and kill (MBC) concentration of C. acnes bacteria less than 0.24 mg ml-1. The particle size of niosomes encapsulate extracts before reducing particle size equal to 1,265 nm. and after reducing particle size in the range of 123 - 219 nm. The skin hydration value after using product for 4 weeks as niosome gel encapsulate extract 2%w w-1 of extract was maximum moisture and minimal water loss.

Grape (Vitis vinifera L.) skin extract reduced levels of SGPT and SGOT and improved the liver tissue structure of Wistar rats (Rattus novergicus) fed a high-cholesterol diet

Background: This study determined the effect of adding grape skin extract (Vitis vinifera L.) to the diet of Wistar rats fed with high cholesterol on levels of SGPT and SGOT and the structure of liver tissue. Methods: This study used 5 groups, namely control (standard diet), P1 (high cholesterol diet), P2 (high cholesterol diet and extract of grape skin as much as 100 mg/200 gBW/day), P3 (high cholesterol diet and extract of grape skin as much as 250 mg/200 gBW/day) and P4 (high cholesterol diet and extract of grape skin 500 mg/200 gBW/day). The study began with the production of extracts, the extracts dosage determination, high cholesterol diet preparation, sample (animals) preparation, extract addition to animals, SGPT and SGOT levels measurement, rat liver histopathology preparation and histopathological features observation, then ended with data analysis. Results: The results showed that administration of grape skin extract in Wistar rats fed a high-cholesterol diet affected the levels of SGPT and SGOT, as well as parenchymatous degeneration of hepatocytes. Administration of grape skin extract as much as 500 mg/200 g BW/day had the lowest mean levels of SGPT and SGOT and the lowest parenchymatous degeneration of hepatocytes. Conclusion: Thus, the extract of grape skin (Vitis vinifera L.) has the potential of hepatoprotector because it contains anthocyanin, which functions as an antioxidant and protects against liver damage.

Rotavirus C Replication in Porcine Intestinal Enteroids Reveals Roles for Cellular Cholesterol and Sialic Acids.

Rotaviruses (RVs) are a significant cause of severe diarrheal illness in infants and young animals, including pigs. Group C rotavirus (RVC) is an emerging pathogen increasingly reported in pigs and humans worldwide, and is currently recognized as the major cause of gastroenteritis in neonatal piglets that results in substantial economic losses to the pork industry. However, little is known about RVC pathogenesis due to the lack of a robust cell culture system, with the exception of the RVC Cowden strain. Here, we evaluated the permissiveness of porcine crypt-derived 3D and 2D intestinal enteroid (PIE) culture systems for RVC infection. Differentiated 3D and 2D PIEs were infected with porcine RVC (PRVC) Cowden G1P[1], PRVC104 G3P[18], and PRVC143 G6P[5] virulent strains, and the virus replication was measured by qRT-PCR. Our results demonstrated that all RVC strains replicated in 2D-PIEs poorly, while 3D-PIEs supported a higher level of replication, suggesting that RVC selectively infects terminally differentiated enterocytes, which were less abundant in the 2D vs. 3D PIE cultures. While cellular receptors for RVC are unknown, target cell surface carbohydrates, including histo-blood-group antigens (HBGAs) and sialic acids (SAs), are believed to play a role in cell attachment/entry. The evaluation of the selective binding of RVCs to different HBGAs revealed that PRVC Cowden G1P[1] replicated to the highest titers in the HBGA-A PIEs, while PRVC104 or PRVC143 achieved the highest titers in the HBGA-H PIEs. Further, contrasting outcomes were observed following sialidase treatment (resulting in terminal SA removal), which significantly enhanced Cowden and RVC143 replication, but inhibited the growth of PRVC104. These observations suggest that different RVC strains may recognize terminal (PRVC104) as well as internal (Cowden and RVC143) SAs on gangliosides. Finally, several cell culture additives, such as diethylaminoethyl (DEAE)-dextran, cholesterol, and bile extract, were tested to establish if they could enhance RVC replication. We observed that only DEAE-dextran significantly enhanced RVC attachment, but it had no effect on RVC replication. Additionally, the depletion of cellular cholesterol by MβCD inhibited Cowden replication, while the restoration of the cellular cholesterol partially reversed the MβCD effects. These results suggest that cellular cholesterol plays an important role in the replication of the PRVC strain tested. Overall, our study has established a novel robust and physiologically relevant system to investigate RVC pathogenesis. We also generated novel, experimentally derived evidence regarding the role of host glycans, DEAE, and cholesterol in RVC replication, which is critical for the development of control strategies.

Dandelion (Taraxacum officinale) seeds extract attenuates hypercholesterolemia in swiss albino mice

ObjectivesThe present search aimed to investigate the effect of dandelion seeds extract to reduce hypercholesterolemia and its hazardous effects on liver. MethodsForty male albino mice were divided into 4 groups, 10 mice per each, the first was the untreated control, the second treated with 400 mg/kg dandelion seed extract, the third treated with 5 mg/kg of cholesterol powder dissolved in oil and the fourth treated with cholesterol and dandelion extract. ResultsIt was showed that dandelion seeds extract reduced the weight of the mice. Cholesterol treatment resulted in highly significant increase of liver enzymes activities, total cholesterol, how-density lipoprotein (HDL) and low-density lipoprotein (LDL) of serum besides to pathological changes as steatosis, inflammation, fibrosis and high pathological score in the liver. Dandelion seeds extraction treatment resulted in marked decrease of liver enzymes activities total cholesterol, HDL and LDL of serum, in addition to hepatic tissue improvement with less steatosis, inflammation, fibrosis and lowered pathological score. ConclusionDandelion seeds extract could reduce hypercholesterolemia in swiss albino mice.

Formulation development & Evaluation of Phospholipids Complex of Euphorbia neriifolia Linn Extract

Phytoconstituents have been utilized as medicines for thousands of years, yet their application is limited owing to major hurdles like deficit lipid solubility, large molecular size and degradation in the gastric environment of gut. To overcome this hurdle, Novel Drug Delivery System (NDDS) proves to be a promising method for formulations containing phytoconstituents. In present work, phyto-phospholipids complex (phytosomes) of Euphorbia neriifolia L. has been prepared to improve the absorption and bioavailability of phytoconstituents. Euphorbia neriifolia L. consists of flavonoids, saponins and polyphenols which possess various medicinal properties. These constituents are hydrophilic in nature and have large molecular size, thus causing poor absorption. Phyto-phospholipids complex of Euphorbia neriifoliahas been prepared by taking Phosphatidylcholine (PC), Cholesterol and Euphorbia neriifolia extract in different ratio and the formulation been optimized to achieve maximum entrapment efficiency and smaller molecular size. The prepared phytosomes has been evaluated by Optical Microscopy, Transmission Electron Microscopy, Dynamic Light Scattering and UV/Vis Spectroscopy for physical appearance, entrapment efficiency, particle size, zeta potential and dissolution rate. The result so obtained indicates the improvement in the absorption rate and bioavailability of the phytoconstituents. Thus, Novel Drug Delivery System (NDDS) possess a great potential in overcoming the challenges of plant based formulations.
 Keywords: Euphorbia neriifolia, Phyto-phospholipids Complex, Novel Drug Delivery System, Phytosomes

Abstract 1151: Inclusion of cholesterol and A549 lipid extract in CLENs improved cellular uptake in adenocarcinoma non-small cell lung cancer target cells

Abstract Purpose:The purpose of this investigation was to determine the optimal amount of cholesterol and A549-derived lipid extract (LE) included in the initial design and development of A-549-cell membrane lipid extracted nanoliposomes (A549-CLENs) for cell targeting studies. Methods and Materials:A549 (ATCC® CCL-185™), used as a positive control and NCI-H1703 (ATCC® CRL-5889™), used as a negative control, and respective growth media were purchased from ATCC (Manassas, VA). A549-GFP (green fluorescent protein) cell line was purchased from AntiCancer, Inc. DOPC, DOTAP, cholesterol, and DSPE-PEG5000 were obtained from Avanti Polar Lipids (Alabastar, AL). Cellular membrane lipids of A549 cells were extracted by modified Bligh & Dyer method. CLENs were fluorescently labeled with rhodamine-B. Particle sizes and zeta potentials were performed using ZetaPALS (Brookhaven Instruments Corporation, Holtsville, NY, USA). For cellular uptake studies cells were seeded at 2.0 x 104 cells in a 48-well plate followed by incubation for 24 hours. A fluorescence microplate reader and/or fluorescence microscopy were used to evaluate cellular uptake as a function of relative amount of cholesterol and A549-lipid extract inclusion in CLENs. Results:In general, an increase in the inclusion of cholesterol reduced the particle size of CLENs. Amount of 10 mol% of cholesterol content in CLENs demonstrated the smallest mean size diameters and a zeta potential value recorded at 243 ± 4.4 nm and 1 ± 2.61 mV, respectively. The finding correlated also with a relatively high degree of cellular uptake of the preparation type for part I of the physicochemical characterization studies. We next maintained 10 mol% cholesterol content consistent in all future preparations and increased the total A-549 lipid extract material at the expense of DOPC. In general, the mean diameter and zeta potential for A549-CLENs were 246 ± 3.8 nm and 20.4 ± 0.84 mV, respectively. Approximately 10-15 mol% of A-549 lipid extract inclusion resulted in a consistently high degree of overall cellular uptake by GFP-A549 and A549 target cells compared to control and the non-target cell population, NCI-H1703. Conclusion: Preparation of A549-CLENs consisting of DOPC/ DOTAP/ Chol/ LE/PEG (65/10/10/10/5) established the greatest increase in cellular uptake and overall selectivity. Future studies will consider antibody conjugation and development of tumor-specific biomarkers for serodiagnostic detection and lung cancer treatment. Citation Format: Maxime Langlois, Guo-Yun Kao, Diana Lasker. Inclusion of cholesterol and A549 lipid extract in CLENs improved cellular uptake in adenocarcinoma non-small cell lung cancer target cells [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1151.

Abstract 246: Glioblastoma cell-modified cell membrane lipid-extracted nanoparticles (CLENs) for imaging and treatment: Influence of magnetic nanoparticle on physiochemical properties of CLENs, cellular uptake, and cytotoxicity

Abstract Background: Glioblastoma multiforme (GBM), located in the cerebral hemispheres of the brain, is the most aggressive form of cancer. Surgery associated with chemotherapy with radiation treatment is the defined way to treat the disease. The delivery of chemotherapeutic agents frequently result in serious side effects. To address this issue we have taken the first steps in the development of CLENs (cell membrane lipid-extracted nanoliposomes) to enhance selective delivery of drugs to the brain. Method: The CLENs used for the experiments were composed of various ratios of DOPC, DOTAP, cholesterol, and lipid extract (LE) material derived from the target cells (i.e., U87-MG), and DPPE-PEG5000 (PEG). Studies include the determination of drug (cisplatin) loading, magnetite (MAG-C) loading in CLENs, quantitative studies for particle size, zeta potential, SQUID (superconducting quantum interference device) measurements, and fluorescence detection to study cellular interactions. Results: The average size for the CLENs fell within the range of 130 to 190 nm. The zeta potential reported for CLENs was negatively-charged. The inclusion of 15mol % LE resulted in the most significant cellular uptake by target cell U87-MG. Overall, the uptake by these cells was less compared to CLENS previously published for other organ tissue environments such as with breast tissue. The inclusion of cationic lipid DOTAP in CLENs did not alter the size distribution; however, the values for zeta potential increased from -8 ± 5 to 12 ± 6 mv. The additional inclusion of DOTAP in CLENs increased cellular uptake approximately 4-fold compared to CLENs without DOTAP (fluorescence intensity (A.U) from 145 ± 52 to 525 ± 49)). The inclusion of DOTAP increased the efficiency of loading Cisplatin in CLENs by approximately 30%. Studies involving the incorporation of MAG-C in U87-MG CLENs are currently underway, as well magnetization studies involving SQUID. Conclusion: All CLENs demonstrated practical physicochemical properties similar to conventional liposomes. The cellular uptake results suggested that cellular uptake of CLENs in vitro, although relatively low overall, varied according to the total LE content. Furthermore, the inclusion of DOTAP in CLENs significantly increased uptake and the efficiency of drug loading. Citation Format: Woongjae Song, Robert Campbell. Glioblastoma cell-modified cell membrane lipid-extracted nanoparticles (CLENs) for imaging and treatment: Influence of magnetic nanoparticle on physiochemical properties of CLENs, cellular uptake, and cytotoxicity [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 246.

ANALYSIS OF APO-B SERUM LEVELS IN BALB/C MICE HYPERCHOLESTEROLEMIC AGAINST TEMULAWAK EXTRACT (Curcuma xanthoriza Roxb)

APO–B Serum levels is the most predictive value for the incidence of atherosclerosis and coronary heart disease. Curcuma xanthorrhiza Roxb contains curcumin, which can be used as an antioxidant, anti–inflammatory and antihypercholesterol. The mechanism of curcumin contained in ginger to reduce cholesterol is due to its function as a cholagoga or bile stimulant. This study aims to determine the effect of temulawak extract on the levels of APO–B Serum in hypercholesterolemia mice. This research were a true experimental study with a post–test only control group design carried out in February 2018. The extraction As much as 25 mice were divided into 5 groups where are group consisted of 5 mice. Positive control group (K+) were treated with high cholesterol feed and water, negative control group (K–) were given standard feed and water, treatment group 1 (P1) were given high cholesterol food and 25mg/kg BW of curcuma extract for 14 days, treatment group 2 (P2) were treated with foods high in cholesterol and 50mg/kg BW of curcuma extract for 14 days and treatment group 3 (P3) treated with high cholesterol and ginger extract 75mg/kg BW for 14 days. Examination of APO–B levels were measured using the spectrophotometric method. Data were analyzed using One–Way Anova. The results showed that the average of APO–B level at (K+) was 209.7 ± 1.02 mg / dL, at (K–) 115.3 ± 1.04 mg / dL, at (P1) 180.4 ± 1.07 mg / dL, at (P2) 147.6 ± 1.12 mg / dL, at (P3) 119.1 ± 1.10 mg / dL. Based on the results of statistical test it was found that there was a significant decrease in APO–B levels with p–value= 0.001 at alpha 0.05 (p <α).

ANTIDIABETIC ACTIVITY OF MORINGA OLEIFERA LAM.

Diabetes mellitus is a clinical syndrome with insufficient insulin secretion, abnormal glucose tolerance, universal, microangiopathia, neuropathia, accentuated atherosclerotic changes. Moringa oleifera Lam (Moringaceae) leaves are emetic and their juice with black pepper is used in headache. The leaves are anthelmintic, aphrodisiac, cures hallucinations, antidiabetic, dry tumors, cough and asthma. Different extracts of Moringa oleifera prepared in different solvents and used in alloxan induced diabetic rats. Total alcohol, Successive alcohol and Successive chloroform significantly decreases the serum cholesterol, serum triglyceride, serum VLDL, serum LDL and Total alcohol extract increases the serum HDL level in alloxan induced diabetic rats. The plant is worth for antidiabetic activity. So this result provides a platform for the drug designers and gives them an opportunity to prepare such herbal formulations which can be used as to treatment on diabetes.

Comparative Study of Protective Effect of Tomato Juice and N-Hexane Extract of Tomato on Blood Lipids and Oxidative Stress in Cholesterol-Fed Rats

Background: Hyperlipidemia is a well-established risk factor in pathogenesis of atherosclerosis and atherosclerosisassociated conditions by enhancing oxidative stress. Dietary supplementation of tomato which is enriched with antioxidants especially like lycopene may therefore be effective in reducing oxidative stress during hyperlipidemic condition. The study evaluates and compares the protective effect of tomato juice and n-Hexane extract of tomato on oxidative stress in hyperlipidemic rats.Methodology: This experimental study was carried out among 42 rats divided in 7 groups. Rats were treated with 0.5% cholesterol (suspension of cholesterol powder in soybean oil) at a dose of 50 mg/ ml once daily through intragastric tube for 8 weeks. In other two groups of rats tomato juice (1 mg/kg) or n-Hexane extract of tomato (1 mg/kg) with 0.5% cholesterol were given orally once daily through intragastric tube for 8 weeks. Serum lipid profile, body weight, plasma malondialdehide (MDA) and erythrocyte reduced glutathione(GSH) levels were measured after 8 weeks in all the groups.Results: Administration of cholesterol caused significant increase (p<0.001) in serum cholesterol, serum triglyceride (TG), serum low-density lipoprotien cholesterol (LDL-C)and significant decrease (p<0.001) in serum high-density lipoprotein cholesterol (HDL-C) which were associated with significant increment (p<0.001) in plasma MDA levels and depletion(p<0.001) in erythrocyte GSH levels. Concomitant treatment of tomato juice or n-Hexane extract of tomato with cholesterol reduced (p<0.001) serum cholesterol, serum LDL-C, body weight and increased serum HDL-C level in cholesterol plus tomato juice treated group (p<0.001) and in cholesterol plus n-Hexane extract of tomato treated group (p<0.05).But serum triglyceride was decreased(p<.05) only in cholesterol plus tomato juice treated group. Simultaneous treatment of tomato juice or n-Hexane extract of tomato with cholesterol decreased (p<0.001) plasma MDA level and increased (p<0.001) erythrocyte GSH level. However, significant differences were noted between the effect of tomato juice and n-Hexane extract of tomato on serum cholesterol (p<0.001), serum triglyceride (p<.05), serum LDL-C (p<0.001), serum HDL-C level(p<0.01) and plasma MDA(p<0.001) and erythrocyte GSH (p<0.001) levels. But no significant difference was noted on body weight.Conclusion: It may be concluded that both tomato juice and n-Hexane extract of tomato offered protection against hyperlipidemia and oxidative stress, but the protection afforded by tomato juice was superior to n-Hexane extract of tomato.Anwer Khan Modern Medical College Journal Vol. 8, No. 1: Jan 2017, P 30-37

Uji Aktivitas Antiproliferasi Formula Liposom Ekstrak Etanol Kunyit (Curcuma domestica) Terhadap Sel Kanker Payudara T47D

Breast cancer is one of deadliest diseases in the world. Turmeric extract was known to have antiproliferative activity. To minimize its toxicity, turmeric extract was encapsulated with liposome, a vesicle lipid bilayer that was functioned as cancer drug carrier in body. This research aimed to determine encapsulation effect of turmeric extract against antiproliferative activity in T47D breast cancer cells through in vitro assay. Liposomes was made using thin layer method and particle size was reduced by extrusion. Materials that was used phosphatidylcholine, cholesterol, and turmeric extract. Optimization of liposomes was made in three formulations with different extract concentrations. The most optimal formulation was formulation with less extract and physical parameters which have smallest precipitates and longest settling time. Evaluation liposome particle size and zeta potential were used DLS, morphology was used TEM, and entrapment efficiency was used dialysis. The most optimal formulation was tested their antiproliferative activity compared with not encapsulated extracts used 3-(4,5-dimethylazole-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method. The result showed that there was antiproliferative activity in encapsulated extracts. IC50 encapsulated extracts was 45.762 μg/ml and IC50 extracts was 36.399 μg/ml. Liposome particle size was below 445 nm. Zeta potential was -7.51 mV. Morphology was LUV and MVV. Entrapment efficiency was 63.80%. It could be concluded that encapsulation of turmeric extract into liposome could reduce its toxicity against cancer cells.

THE EFFECT OF MUNDU FRUIT (Garcinia dulcis (ROXB.) KURZ) METHANOL EXTRACT ON LYPHOPROTEIN PROFILE AND TRYGLISERIDE WHITE RAT (Rattus norvegicus L)

Garcinia dulcis (Roxb.) Kurz) (Guttiferae) is a tropical fruit tree native to Indonesia and its known as an Asian medicinal plant used in folk medicines. In ethno pharmacology, Garcinia dulcis has been used in traditional medicine, such as anti-inflammatory, anti-tumor, anti-malaria and anti-bacterial. The scientific studies of biological activity of Garcinia dulcis on lipide profile and trigliseride are relatively measly. Therefore, the objectives of this research were to know the effect of methanol extract of Garcinia dulcis fruit on total cholestrol, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol and triglyseride rat hyperlipidemic. These studies using Garcinia dulcis fruit from Banyuasin, South Sumatra. In this observation, 30 male rat from Laboratory and Research Institute for Integrated Testing, Gadjah Mada University, 2 months old were used and divided into five groups randomly. The first group was given high cholesterol and lipid diet, as a normal group. The second group was given high cholesterol, lipid diet and simvastatin 3,6 mg/kg bow/day dosage, as a simvastatin group. The third, fourth and fifth groups was given high cholesterol, lipid diet and methanol extract 1.8; 2.7 and 3.6 mg/kg bow/day dosage. Total cholestrol, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol and triglyseride contains using spectrophotometry method, were observed 1 and 30 days. The differences among groups were analyzed using Anova followed by LSD to locate the differences. LSD analysis showed that Garcinia dulcis methanol extract treatment doses of 3.6 mg/kg bw and simvastatin lowers total cholesterol levels highest in the amount of 32.17 mg/dL and 24.1 mg/dL, significantly different compared with the treatment dose of 1.8 mg/kg bw. Garcinia dulcis fruit methanol extract treatment doses of 3.6 mg/kg bw and simvastatin lowers triglyceride levels by 7.22 mg/dL and 24.58 mg/dL, significantly different compared with the treatment dose of 1.8 mg/kg bw and controls. Garcinia dulcis fruit extract dose of 3.6 mg/dL lower low density lipoprotein-cholesterol highest of 16.45 mg/dL. Garcinia dulcis fruit methanol extract dose of 3.6 mg/kg bw was also able to increase the levels of high density lipoprotein-cholesterol by 5.62 mg/dL. The results showed that the fruit of Garcinia dulcis contain compounds that have potential as agents antihiperlipidemia and presumably through inhibition of the enzyme HMG-CoA reductase

Antidiabetic and antihyperlipidemic activity of hydroalcoholic extract of Withania coagulans Dunal dried fruit in experimental rat models

Objective:Evaluation of antidiabetic potential of the hydroalcoholic extract of Withania coagulans Dunal dried fruit (WCDF) alone and in combination with glipizide, in streptozotocin-induced diabetes, and evaluation of possible antihyperlipidemic activity of the same extract in high-cholesterol diet-induced hyperlipidemia, in albino rats.Materials and Methods:Experimental diabetes was induced in 30 albino rats with intraperitoneal injection of streptozotocin (55 mg/kg). The rats were divided into five groups receiving the following treatments orally for 4 weeks: Vehicle, glipizide (2.5 mg/kg), WCDF extract (1000 mg/kg), WCDF extract (1000 mg/kg) plus glipizide (1 mg/kg) and WCDF extract (1000 mg/kg) plus glipizide (2.5 mg/kg). Fasting and postprandial blood glucose levels were measured every week for 4 weeks. Endocrine pancreas histopathology was done at the end. In a separate set of experiment, five groups of six albino rats each, received orally for 4 weeks, vehicle, cholesterol (25 mg/kg/day), cholesterol (25 mg/kg/day) plus atorvastatin (7.2 mg/kg/day), cholesterol (25 mg/kg/day) plus WCDF extract (1000 mg/kg/day) and no treatment, respectively. Estimation of serum lipid profile and liver histopathology was done at the end of 4 weeks.Statistical Analysis:Between-group and within-group comparisons were respectively done by analysis of variance (ANOVA) and repeated measures ANOVA, followed by post hoc Tukey's test, with a significance level of P < 0.05.Results and Conclusions:The 4-week treatment with WCDF extract significantly reversed hyperglycemia in streptozotocin-induced diabetes that was comparable to glipizide. When combined with glipizide (2.5 mg/kg), WCDF extract produced a synergistic antihyperglycemic effect as well as improvement in pancreatic histopathology. Moreover, hydroalcoholic extract of WCDF was effective and comparable to atorvastatin in controlling the high-cholesterol diet-induced hyperlipidemia in rats.

Enhanced production of cell-bound cholesterol oxidase from Rhodococcus sp. NCIM 2891 by the statistical method

Classical (one-variable-at-a-time) and statistical methods (Plackett-Burman and Central composite design) were used to optimise growth medium for the production of cholesterol oxidase (COX) from Rhodococcus sp. NCIM 2891. COX activities from the classically and statistically optimised media were 0.75 and 3.25 U/ml, respectively. The statistically optimised medium had 4.33- and 9.7-fold higher enzymatic activity than the classically optimised and un-optimised basal medium, respectively. The ratio of enzyme production to cell growth rate was 29-fold higher in our statistically optimised medium than in the basal medium, indicating that the enzyme production could be classified as mixed type of growth. Cell-bound COX accounted for 90.68 ± 2 % of the total enzymatic activity of the growth medium. Interactions between the COX-inducing substrate cholesterol and medium growth substrates yeast extract and (NH4)2HPO4 significantly enhanced the production of cell-bound COX. Our results validate the statistical approach as a potential technique for achieving the large-scale production of cell-bound COX from Rhodococcus sp. NCIM 2891.

남해군 특용작물 열수 추출분말이 콜테스테롤 급이 흰쥐의 혈청 및 장기 중 지질성분에 미치는 영향

남해군의 특용작물인 울금, 백년초, 알로에 및 아스파라거스가 혈청 지질 개선에 미치는 영향을 비교 분석하고자 1%의 콜레스테롤 첨가식이로 유발된 고콜레스테롤혈증 흰쥐를 이용하여 정상군, 고콜레스테롤 투여군을 대조군으로 하고, 특용작물 열수추출물 동결건조한 분말 1% 첨가 급이군을 실험군으로 하여 4주간 급이 후 실험하였다. 식이 섭취량과 체중증가는 실험군에 따른 유의차는 없었으나, 식이효율은 울금 추출분말 1% 급이군이 유의적으로 낮았다. 혈당은 실험군이 대조군에 비해 6.18~23.44% 정도 감소하였으며, 혈중 총 지질은 백년초 추출분말을 1% 급이군에서 374.99<TEX>${\pm}$</TEX>1.54 mg/dL로 실험군중에 가장 낮은 함량이었다. 총 콜레스테롤은 알로에 추출분말 첨가 급이군에서 가장 낮은 함량이었으며, 이는 정상군과 유의차가 없었다. 혈중 중성지질의 함량 또한 대조구에 비해 특용작물 추출분말 첨가군에서 낮았는데 아스파라거스 추출분말 첨가 급이군에서 50.00<TEX>${\pm}$</TEX>6.29 mg/dL로 가장 낮은 함량이었다. HDL-콜레스테롤은 울금 추출분말 급이군이 32.55<TEX>${\pm}$</TEX>6.16 mg/dL로 여타 실험군에 비해 유의적으로 높았으며, 정상군과 유의차가 없었다. LDL-콜레스테롤은 알로에 추출분말 급이군이 대조군에 비해 약 44% 더 낮았으며, VLDL-콜레스테롤은 아스파라거스 급이군에서 유의적으로 낮아 7.77<TEX>${\pm}$</TEX>0.72 mg/dL로 정상군 보다 29%나 더 낮았다. AI와 CRF는 울금과 알로에 추출분말 첨가군이 가장 낮았으며, 혈중 TBARS 함량과 DPPH radical 소거능은 백년초 추출분말 급이군에서 가장 효과적이었다. 간장에 함유되어 있는 총 지질 및 콜레스테롤 함량은 알로에 추출분말 급이군에서 가장 낮아 각각 134.54<TEX>${\pm}$</TEX>5.62 mg/g과 12.25<TEX>${\pm}$</TEX>3.07 mg/g 이었으며, TBARS 함량도 알로에 추출분말 첨가군이 가장 낮았다. This study was performed to investigate the effect of water extracts from Namhae special crops (NSC) on improved serum lipid composition using rats fed a 1% cholesterol diet for 4 weeks. Male Wister rats (200-210 g) were divided into six groups: Normal cholesterol diet group (Normal), 1% cholesterol diet group (Control), 1% cholesterol and NSC water extract powder supplemented groups, including, turmeric (Tu-EP), cactus (Ba-EP), aloe vera (Al-EP) and asparagus (As-EP). No significant differences were observed for food intake or body weight gain between the control and experimental groups. However, food efficiency ratio (FER) was the lowest in the As-EP group. The serum levels of total cholesterol and triglycerides in the NSC water powder extract supplement groups were lower than those in the control group. The serum high density lipoprotein (HDL)-cholesterol content in the Tu-EP group was higher than that in the other experimental groups. Very low density lipoprotein (VLDL)-cholesterol content in the As-EP group was similar to that in normal group. Furthermore, the VLDL content in the Al-EP group was lower than that in the normal group. Serum antioxidant activity by TBARS level and DPPH radical scavenging were significantly higher in the Ba-EP group than that in the control group. Hepatic total cholesterol and lipid content in the Al-EP group decreased significantly compared to that in the control group. These results suggest that the NSC water extract may reduce serum cholesterol and prevent oxidative stress by stimulating antioxidative systems in rats fed a 1% cholesterol diet.

Determination of cholesterol levels in eggs of local and hybrid chicken

Cholesterol, the most abundant sterol in animal tissue was extracted from different egg yolk samples using acetone followed by methanol after filtration and evaporation respectively. The cholesterol extract was coupled with jerk reagent to enhance absorption at visible light region. The cholesterol was then quantified by spectrophotometry using a UV-double beam spectrophotometer. It was found that the hybrid egg had the least value of cholesterol hence is safer for consumption based on the cholesterol content.Keywords: cholesterol, egg, hybrid chicken, egg yokeJournal of Pharmaceutical and Allied Sciences, Vol. 7 No. 1 (2010)

3H]Epibatidine Photolabels Non-equivalent Amino Acids in the Agonist Binding Site of Torpedo and α4β2 Nicotinic Acetylcholine Receptors

Nicotinic acetylcholine receptor (nAChR) agonists, such as epibatidine and its molecular derivatives, are potential therapeutic agents for a variety of neurological disorders. In order to identify determinants for subtype-selective agonist binding, it is important to determine whether an agonist binds in a common orientation in different nAChR subtypes. To compare the mode of binding of epibatidine in a muscle and a neuronal nAChR, we photolabeled Torpedo alpha(2)betagammadelta and expressed human alpha4beta2 nAChRs with [(3)H]epibatidine and identified by Edman degradation the photolabeled amino acids. Irradiation at 254 nm resulted in photolabeling of alphaTyr(198) in agonist binding site Segment C of the principal (+) face in both alpha subunits and of gammaLeu(109) and gammaTyr(117) in Segment E of the complementary (-) face, with no labeling detected in the delta subunit. For affinity-purified alpha4beta2 nAChRs, [(3)H]epibatidine photolabeled alpha4Tyr(195) (equivalent to Torpedo alphaTyr(190)) in Segment C as well as beta2Val(111) and beta2Ser(113) in Segment E (equivalent to Torpedo gammaLeu(109) and gammaTyr(111), respectively). Consideration of the location of the photolabeled amino acids in homology models of the nAChRs based upon the acetylcholine-binding protein structure and the results of ligand docking simulations suggests that epibatidine binds in a single preferred orientation within the alpha-gamma transmitter binding site, whereas it binds in two distinct orientations in the alpha4beta2 nAChR.

효소처리한 함초(Salicornia herbacea) 열수추출물이 고콜레스테롤 식이 흰쥐의 지질대사에 미치는 영향

본 연구에서는 고콜레스테롤 식이 흰쥐에서 효소 처리하여 제조된 함초 열수추출물의 공급이 혈청 및 간조직의 지질대사에 미치는 영향을 관찰하였다. 각 실험군의 100 g당 간 조직의 무게를 비교한 결과 정상식이 대조군과 정상식이에 함초 추출물을 공급한 군은 유의적인 차이가 없었으며, 고콜레스테롤군들에서 정상식이 대조군에 비해 유의적으로 증가하였다. 간조직 중성지질 함량은 고콜레스테롤군들에서 정상식이 대조군에 비해 유의적으로 증가하였으며, 고콜레스테롤식이에 함초 추출물 2% 및 4%를 공급한 군들에서 고콜레스테롤식이 대조군보다 유의적으로 감소되었으며, 간조직 총지질 함량 또한 중성지질과 같은 경향을 나타내었다. 혈청 총콜레스테롤 함량은 고콜레스테롤 식이대조군에서 정상식이 대조군에 비해 유의적으로 증가하였으나, 고콜레스테롤식이에 함초 추출물을 공급함으로써 총콜레스테롤 함량이 다소 감소하였다. 간조직 총콜레스테롤의 함량은 혈청 총 콜레스테롤 함량과 유사한 경향을 나타내었으며, 고콜레스테롤식이에 함초 추출물 2% 및 4%를 공급한 군들에서 고콜레스테롤식이 대조군에 비해 콜레스테롤의 함량이 유의적으로 감소되었다. 혈청 HDL-콜레스테롤 함량은 고콜레스테롤군들에서 정상식이 대조군에 비해 유의적으로 감소하였으며, 고콜레스테롤식이에 함초 추출물을 공급한 군들에서 다소 증가하였다. 혈청 LDL-콜레스테롤 함량은 고콜레스테롤식이 대조군이 정상식이 대조군에 비해 유의적으로 증가하였으나, 고콜레스테롤식이에 함초 추출물 4%를 공급한 군에서 고콜레스테롤식이 대조군에 비해 유의적으로 감소되었다. 따라서, 고콜레스테롤 식이 흰쥐에서 효소처리한 함초 열수추출물의 공급은 혈청 및 간조직의 총지질, 중성지질, 총콜레스테롤, LDL-콜레스테롤 및 HDL-콜레스테롤 등의 지질함량을 조절함으로써 지질대사 개선 효과가 있음을 알 수 있었다.

The water-soluble extract of chicory reduces cholesterol uptake in gut-perfused rats

The direct action of soluble fiber analogs (chicory water-soluble extract and inulin) on the intestinal absorption of cholesterol was investigated in gut-perfused rats. After equilibrium, both jejunal and ileal segments were simultaneously perfused with an isotonic electrolyte solution (pH 7.4) containing cholesterol (2 mmol/L) and chicory water-soluble extract (chicory extract) or inulin (10 g/L). Each test or control solution was perfused in random sequence, with perfusion times of 30 min. Chicory extract significantly decreased cholesterol absorption by 30% (P < 0.05) in the jejunum and by 41% (P < 0.05) in the ileum, compared with control. Addition of inulin resulted in a significant reduction of cholesterol absorption from jejunum by 39% (P < 0.05) and from ileum by 51% (P < 0.05). The observed changes in cholesterol absorptions caused by chicory extract or inulin were reversible by switching to a fiber-free (control) perfusate. The reduction in intestinal absorption of cholesterol observed after perfusion of chicory extract or inulin may be due to its increased viscosity of intestinal contents.

Maximization of cholesterol oxidase production by Rhodococcus equi no. 23 By using response surface methodology.

Medium optimization for the production of cholesterol oxidase (EC 1. 1.3.6) by Rhodococcus equi no. 23 was investigated by using response surface methodology and a central composite design. Results revealed that cholesterol and yeast extract had positive effects and the interaction between any two of three factors had no significant effect on cholesterol oxidase production. The optimized medium was the basal medium with the addition of 2.30 g/l cholesterol, 8.18 g/l yeast extract and 4.10 ml/l Tween 80. The peak cholesterol oxidase production (0.242 unit/ml) after 60-72 h cultivation was approx. 4-fold that in control medium.