Enzymatic Kit Research Articles

A new osmium-polymer modified screen-printed graphene electrode for fructose detection

This paper describes the development and performance of the first fructose biosensor based on a commercial screen-printed graphene electrode (SPGE). The electrode was modified with an osmium-polymer, which allowed the efficient wiring of the enzyme fructose dehydrogenase (FDH). The immobilization of both osmium-polymer and FDH was realized in an easy way. Aliquots of 10μL Os-polymer and 10μL FDH were thoroughly mixed with poly(ethylene glycol) (400) diglycidyl ether (PEDGE) and deposited on the electrode surface and left there to dry overnight. The biosensor exhibits a detection limit of 0.8μM, a linear range between 0.1 and 8mM, high sensitivity to fructose (2.15μAcm−2/mM), good reproducibility (RSD=1.9%), fast response time (3s) and a stability of 2 months when stored in the freezer.The proposed fructose biosensor was tested in real food samples and validated with a commercial spectrophotometric enzymatic kit. No significant interference was observed with the proposed biosensor.

Detection of Coliforms in Drinking Water Using Skin Patches: A Rapid, Reliable Method that Does Not Require an External Energy Source

The detection of coliforms requires incubation in a laboratory, generally powered using electricity. In many parts of the developing world, however, external energy sources such as electricity are not readily available. To develop a fast, reliable method for detecting coliforms in water without an external energy source, we assessed the efficacy of six test kits for the identification of coliforms in water samples. To assess the possibility of using body temperature as the sole source of heat for incubation, bacterial samples were then mixed with the enzymatic test kit reagent and attached to the human body surface using a patch system. The patches were attached to the bodies of volunteers for 24 hours and the practicality and accuracy of the patches were assessed. Coliforms were detected within 24 hours in all patches. This innovation will facilitate the testing of water quality by researchers and by economically disadvantaged people without electricity.

Comparison of DNA Extraction Methods for Human Oral Microbiome Research

The oral micro biome is highly diverse and its composition is associated with oral disease and potentially diseases at other sites. Our objective is to evaluate DNA extraction methods potentially suitable for population-based investigations on the oral human microbiome and disease risk. Six commonly used microbial DNA extraction kits, employing either enzymatic methods or mechanical bead beating for cell lysis, were evaluated for the following aspects total DNA yield and quality and 16s rRNA DNA product and representation of microbial diversity. All analyses were carried out using a pooled and homogenized sample from one study subject. 16s rRNA gene sequence data were processed using the QIIME pipeline. One way ANOVA and Kruskal-Wallis tests were used to compare the different DNA extraction methods. We found that enzymatic extraction kits produced higher human genomic DNA, compared with mechanical extraction kits, however, phylogenic diversity in oral microbiome community structure from 16s rRNA gene sequence reads revealed no important differences between kit types. Enzymatic and mechanical bead beating kits provide alternative approaches for DNA extraction of oral microbiome DNA from oral wash samples. Greater total DNA yields are found in enzymatic approaches but microbial diversity can be similarly well characterized by either enzymatic or mechanical bead beating approaches. Original Research Article British Journal of Medicine & Medical Research, 4(10): 1980-1991, 2014 1981

Rapid monitoring of glycerol in fermentation growth media: Facilitating crude glycerol bioprocess development

Recently, the need for crude glycerol valorisation from the biodiesel industry has generated many studies for practical and economic applications. Amongst them, fermentations based on glycerol media for the production of high value metabolites are prominent applications. This has generated a need to develop analytical techniques which allow fast and simple glycerol monitoring during fermentation. The methodology should be fast and inexpensive to be adopted in research, as well as in industrial applications. In this study three different methods were analysed and compared: two common methodologies based on liquid chromatography and enzymatic kits, and the new method based on a DotBlot assay coupled with image analysis. The new methodology is faster and cheaper than the other conventional methods, with comparable performance. Good linearity, precision and accuracy were achieved in the lower range (10 or 15 g/L to depletion), the most common range of glycerol concentrations to monitor fermentations in terms of growth kinetics.

Bioactive Amines: Aspects of Quality and Safety in Food

Presented in several types of food, bioactive amines are described as organic bases of low molecular weight. They have vasoactive, psychoactive and toxicological characteristics and constitute a potential health risk. The concentration of amines formed in foods depends on the type of microorganisms present, the action of decarboxylase enzymes produced by microorganisms on specific amino acids and favorable conditions for enzymatic activity. The presence of these chemical metabolites has been suggested as a quality indicator in routine analyzes for food production and marketing monitoring. The detection of bioactive amines can be performed by chromatographic methods, fluorometric and enzymatic kits. Bioactive amine formation can be prevented mainly through the adoption of good manufacturing practices, but the industry can also use other methods such as temperature control in the production chain, modified atmosphere packaging and food irradiation. This review aims to address the formation of bioactive amines in foods, emphasizing the formation and classification of these metabolites, aspects related to health, acceptable limits, detection methods and control methods used in the industry to ensure food safety and quality. The success of this approach is linked to the importance of bioactive amines as quality indicators, as well as the discussion on the development of methodologies for determining these substances and discussion of acceptable parameters in food.

Correlation of serum paraoxonase activities in known cases of 130 elderly hypertensive South Asian aged 56–64 years – a hospital based study

ObjectiveTo evaluate paraoxonase activity, antioxidant status and lipid peroxidation in hypertensive participants and to address the hypothesis that oxidative modifications of lipids due to hypertension can cause changes in serum paraoxonase activities. MethodsThe serum paraoxonase activities, antioxidants and lipid peroxidation were determined in 130 hypertensive participants and 130 age-sexes matched normotensive healthy volunteers served as control. Serum paraoxonase activities were measured by enzymatic kit. The glutathione peroxidase, superoxide dismutase and catalase activity were determined by standard methods. Malondialdehyde was measured by thiobarbituric acid reaction. Conjugated diene level was measured by Recknagel and Glende method. Serum uric acid, total bilirubin, serum albumin, serum ascorbic acid and lipid profile were analyzed by standard methods. ResultsTotal cholesterol, triglycerides, low-density lipoprotein cholesterol were significantly higher and high-density lipoprotein cholesterol were significantly lower in hypertensive patients when compared to normotensive healthy controls. The superoxide dismutase, glutathione peroxidase and catalase were significantly lower in hypertensive when compared with normotensive. Similar findings were observed in the levels of albumin, uric acid, bilirubin and ascorbic acid when hypertensives were compared with normotensive. The oxidative stress indicators namely malondialdehyde and conjugated diene were significantly higher and paraoxonase activity were significantly lower in hypertensive. ConclusionsOur study concludes that paraoxonase activities are bound to alter in hypertension which is caused due to interplay of several confounding factors namely oxidative stress, increased oxidized low-density lipoprotein and depletion of antioxidants.

Effect of Natural Product Extracts on Lipoxygenase, Cyclooxygenase, and Protein Tyrosine Phosphatase 1β

The purpose of this research project is to examine the inhibition of enzymes linked to inflammation and associated diseases by plants traditionally as anti-inflammatory medicines: Tussilago farfara, Grindelia squarrosa, and Uritca dioica. Arachidonic acid is metabolized in the body through two main metabolic pathways with the enzymes: cyclooxygenases (COX) and lipoxygenases (LOX). Elevated levels of prostanoids and leukotrienes, products of the two respective pathways, have been linked to inflammatory diseases. Finding a dual inhibitor of COX and LOX is promising in preventing the inflammation and diseases that are linked to the overproduction of both pathways while minimizing the side effects associated with inhibition of individual pathways. Furthermore, the enzyme protein tyrosine phosphatase 1β (PTP1β) is linked between inflammation and metabolic disease through the leptin receptor-associated Janus Kinase (JAK). PTP1β is a negative regulator of insulin and leptin receptors thus being explored as a possible therapeutic for type II diabetes and obesity. Crude methanolic extracts of Tussilago farfara, Grindelia squarrosa, and Urtica dioica were taken to approximate the plants' components released in the body upon consumption. The bioactivities of the standardized extracts were then determined using enzymatic assay kits for COX I and II, LOX, and PTP1β. T. farfara is the strongest LOX inhibitor at 60% inhibition of 15-LOX, followed by U. dioica and G. squarrosa at 45% and 39%, respectively. Studies for COX I and II as well as PTP1β inhibition are ongoing, and the results will be presented.

Variation in Lipid Profile during Different Interval of Gestation Period

Just after conception, there is a highly increased demand of energy and carbon for various metabolic or anabolic pathways rather to furnish the molecular requirement of foetus and mother as well. To achieve the target, there are lot of changes that occur at every level belonging to anatomy, physiology and biochemistry. An inclined physiological factor reflects in the biochemical parameter of the subject. Lipid profile is one of them. The lipid profile is major element and prerequisite to atherosclerosis and may cause unconsciousness. To save mankind from this problem, it is planned to observe and analyse the changes in the lipid profile during various trimesters of gestation period. For this research work, 80 subjects of 30-40 age group were selected against 50 subjects of the same age group as a control. Overnight fasting at least 10 h is allowed before taking the sample. Total cholesterol, HDL, LDL, VLDL, triglyceride were estimated by the enzymatic colorimetric kit method in Divya Jyoti Institute of Medical Sciences and Research, Niwari Road, Modinagar, Ghaziabad, UP, India. Total cholesterol, LDL, VLDL, triglyceride increased while HDL decreased from the first to third trimester in pregnant women. This may change the lipid profile, associated with length of gestation period are observed disturbed in each trimester and this explains that variation in the lipid profile is somehow associated with the gestation period. In brief, there is a chance of elevation for coronary heart disease and other ailments concerned with mother/foetal health. This is suggestive that the routine lipid profile analysis is necessary to prevent the risk during the antenatal period. Studies are required further to observe whether pregnancy affects their life from coronary vascular disease in the later stage or not.

Apolipoprotein A-IV Reduces Hepatic Gluconeogenesis through Nuclear Receptor NR1D1

We showed recently that apoA-IV improves glucose homeostasis by enhancing pancreatic insulin secretion in the presence of elevated levels of glucose. Therefore, examined whether apolipoprotein A-IV (apoA-IV) also regulates glucose metabolism through the suppression of hepatic gluconeogenesis. The ability of apoA-IV to lower gluconeogenic gene expression and glucose production was measured in apoA-IV(-/-) and wild-type mice and primary mouse hepatocytes. The transcriptional regulation of Glc-6-Pase and phosphoenolpyruvate carboxykinase (PEPCK) by apoA-IV was determined by luciferase activity assay. Using bacterial two-hybrid library screening, NR1D1 was identified as a putative apoA-IV-binding protein. The colocalization and interaction between apoA-IV and NR1D1 were confirmed by immunofluorescence, in situ proximity ligation assay, and coimmunoprecipitation. Enhanced recruitment of NR1D1 and activity by apoA-IV to Glc-6-Pase promoter was verified with ChIP and a luciferase assay. Down-regulation of apoA-IV on gluconeogenic genes is mediated through NR1D1, as illustrated in cells with NR1D1 knockdown by siRNA. We found that apoA-IV suppresses the expression of PEPCK and Glc-6-Pase in hepatocytes; decreases hepatic glucose production; binds and activates nuclear receptor NR1D1 and stimulates NR1D1 expression; in cells lacking NR1D1, fails to inhibit PEPCK and Glc-6-Pase gene expression; and stimulates higher hepatic glucose production and higher gluconeogenic gene expression in apoA-IV(-/-) mice. We conclude that apoA-IV inhibits hepatic gluconeogenesis by decreasing Glc-6-Pase and PEPCK gene expression through NR1D1. This novel regulatory pathway connects an influx of energy as fat from the gut (and subsequent apoA-IV secretion) with inhibition of hepatic glucose production.

Novel N-substituted aminobenzamide scaffold derivatives targeting the dipeptidyl peptidase-IV enzyme

BackgroundThe dipeptidyl peptidase-IV (DPP-IV) enzyme is considered a pivotal target for controlling normal blood sugar levels in the body. Incretins secreted in response to ingestion of meals enhance insulin release to the blood, and DPP-IV inactivates these incretins within a short period and stops their action. Inhibition of this enzyme escalates the action of incretins and induces more insulin to achieve better glucose control in diabetic patients. Thus, inhibition of this enzyme will lead to better control of blood sugar levels.MethodsIn this study, computer-aided drug design was used to help establish a novel N-substituted aminobenzamide scaffold as a potential inhibitor of DPP-IV. CDOCKER software available from Discovery Studio 3.5 was used to evaluate a series of designed compounds and assess their mode of binding to the active site of the DPP-IV enzyme. The designed compounds were synthesized and tested against a DPP-IV enzyme kit provided by Enzo Life Sciences. The synthesized compounds were characterized using proton and carbon nuclear magnetic resonance, mass spectrometry, infrared spectroscopy, and determination of melting point.ResultsSixty-nine novel compounds having an N-aminobenzamide scaffold were prepared, with full characterization. Ten of these compounds showed more in vitro activity against DPP-IV than the reference compounds, with the most active compounds scoring 38% activity at 100 μM concentration.ConclusionThe N-aminobenzamide scaffold was shown in this study to be a valid scaffold for inhibiting the DPP-IV enzyme. Continuing work could unravel more active compounds possessing the same scaffold.

가시오가피와 탐라오가피의 산지별 및 부위별 Eleutheroside B, E 및 β-Glucan 함량 분석

The present study was performed to quantitatively analyze eleutherosides (B and E) and β-glucan in different plant parts of three cultivars (Chungnam, Gangwon, and Jeju) of Acanthopanax senticosus and Acanthopanax koreanum using HPLC and a commercial enzyme kit. Our results showed high linearity in the calibration curves as the coefficients of correlation (R2) were 0.998 (eleutheroside B) and 0.999 (eleutheroside E), respectively. Eleutheroside B and E were found in stem extracts of A. koreanum cultivated in Jeju (1,122 μg/g, eleutheroside B) and A. senticosus cultivated in Chungnam (2,536 μg/g, eleutheroside E), respectively. However, eleutheroside B was not detected in any part of A. senticosus cultivated in Chungnam. For β-glucan contents, stems of A. senticosus and A. koreanum showed higher than other parts. Furthermore, the β-glucan content in stems of A. koreanum cultivated in Gangwon was significantly higher than in those of other cultivars. These results show that the contents of eleutheroside B, E, and β-glucan were higher in stem extracts of A. senticosus and A. koreanum than other parts. Moreover, our results suggest that the contents of eleutheroside B, E, and β-glucan in A. senticosus and A. koreanum are influenced by cultivation area and the selected part.

Use of Polyhydroxybutyrate and Ethyl Cellulose for Coating of Urea Granules

Fertilizers contain essential nutrients for agricultural growth and development. However, most nitrogen fertilizers are substances with high solubility of ions and are very susceptible to leaching and volatilization. To minimize these losses, an alternative is the creation of a physical barrier around granules. One way is to coat granules with polymers. In the present work urea granules were coated with polyhydroxybutyrate and ethyl cellulose in various conditions in the presence of emulsifiers. The original granules and the final products were characterized by scanning electron microscopy, Fourier transform infrared spectroscopy and thermogravimetry, to evaluate the surface morphology, the interaction between the granules and the coating, and the rates of mass change. The rates of urea release in distilled water were measured with a commercial enzyme kit. It is shown that those polymers are effective for coating of granules, leading to reduction of rates of urea dissolution in water.

Effects of small interfering RNA-mediated hepatic glucagon receptor inhibition on lipid metabolism in db/db mice

Hepatic glucose overproduction is a major characteristic of type 2 diabetes. Because glucagon is a key regulator for glucose homeostasis, antagonizing the glucagon receptor (GCGR) is a possible therapeutic strategy for the treatment of diabetes mellitus. To study the effect of hepatic GCGR inhibition on the regulation of lipid metabolism, we generated siRNA-mediated GCGR knockdown (si-GCGR) in the db/db mouse. The hepatic knockdown of GCGR markedly reduced plasma glucose levels; however, total plasma cholesterol was increased. The detailed lipid analysis showed an increase in the LDL fraction, and no change in VLDL HDL fractions. Further studies showed that the increase in LDL was the result of over-expression of hepatic lipogenic genes and elevated de novo lipid synthesis. Inhibition of hepatic glucagon signaling via siRNA-mediated GCGR knockdown had an effect on both glucose and lipid metabolism in db/db mice.

A simple method for the determination of the cholesterol esterase activity.

The proposed method determines the activity of cholesterol esterase (CEH) and takes advantage of its ability to catalyze the hydrolysis of cholesterol esters naturally present in human serum. The assay is based on Allain's method of spectrophotometric determination of cholesterol by means of cholesterol oxidase, peroxidase, but using 3,5-dichloro-dihydroxybenzenesulfonic acid (DHBS) as phenolic chromogen and human serum as a source of substrate for the CEH as a novelty. Furthermore, it is characterized by low costs and high precision. It can be employed to control the activity of CE preparations used for the preparation of enzymatic kits for the determination of cholesterol or for screening of potential bacterial enzyme producers.

RETRACTED: Neuroprotective effect of Liuwei Dihuang decoction on cognition deficits of diabetic encephalopathy in streptozotocin-induced diabetic rat

Liuwei Dihuang decoction (LWDHD) is a well-known prescription of traditional Chinese medicine (TCM) and consists of six crude drugs including Rehmannia glutinosa Libosch. (family: Scrophulariaceae), Cornus officinalis Sieb. (family: Cornaceae), Dioscorea oppositifolia L. (family: Dioscoreaceae), Paoenia ostii (family: Paeoniaceae), Alisma orientale (G. Samuelsson) Juz (family: Alismataceae) and Poria cocos (Schw.) Wolf (family: Polyporaceae). It has been used for the treatment of "Kidney-Yin" deficiency syndrome in clinic in China for a long time. Recent studies found that LWDHD had a potential benefit for the treatment of diabetic complications. The aim of the present study is to investigate the neuroprotective effect of LWDHD on memory and cognition deficits in streptozotocin (STZ)-induced diabetic encephalopathy (DE) rats. Adult male Sprague Dawley (SD) rats were fed with high-glucose-fat diet for 50 days and then received an intraperitoneal injection of STZ (40 mg/kg) to induce DE model. Morris water maze test was used to evaluate the memory and cognition capability of DE rats. Choline acetyltransferase (ChAT), acetylcholinesterase (AChE), Na(+)-K(+)-ATP enzyme, iNOS and GSH kits were used to determine their activities or content in hippocampus. TUNEL staining, immunohistochemistry and Congo red staining were conducted to evaluate the apoptosis, caspase-3 protein expression, insulin-like growth factors 1 (IGF-1) and brain derived neurophic factor (BDNF) expressions, as well as Aβ deposition. The treatment with LWDHD (1 and 2g/kg, p.o., once daily, 30 days) could significantly reduce the escape latency time and path length, and obviously enhance the spent time in the target quadrant and platform crossings in Morris water maze test compared with model group (P<0.05, P<0.01). LWDHD could also significantly decrease the level of fasting blood glucose, increase Na(+)-K(+)-ATP enzyme and ChAT activities, enhance remarkedly GSH level while decrease significantly AChE and iNOS activities in hippocampus (P<0.05, P<0.01). Furthermore, TUNEL staining, Congo red staining and immunohistochemistry showed that LWDHD significantly improved the expressions of IGF-1 and BDNF, attenuated the neural apoptosis, overexpression of caspase-3 and Aβ deposition in the hippocampus and cerebral cortex of STZ-induced DE rats (P<0.01). Our findings suggested that LWDHD had a neuroprotective effect on DE rats. LWDHD may be of benefit in the treatment of DE.

Analytical performance and accuracy verification of four domestic and two imported enzymatic creatinine assay kits

Objective To assessed the analytical performance of six commercial enzymatic creatinine (Cr) reagents and compared the accuracy among them.Four reagents came from domestic manufactures (Maker Biotechnology,Beijing Strong Biotechnologies,Biosino Bio-technology & Science,BaiDing Biological Engineering,labeled as A,B,C,D respectively) and the other two reagents came from oversea manufactures (Sekisui Medical Technology,Japan and Wako Pure Chemical Industries,Japan,labeled as E,F respectively). Methods Performance verification test.70 serum samples were obtained from chronic kidney disease outpatients and medical center healthy volunteers in Peking Union Medical College Hospital from May 2010.Among 30 patients ureh CKD (15 moles,15 females) aged 18 to 80 years old;40 healthy volunteers (20 males,20 females) aged 20 to 60 years old.To evaluate the within-run imprecision and total imprecision according to the CLSI EP15-A protocol,two level control materials were measured with six reagents in five days,with a frequency of four times a day.To verify the linear ranges according to the EP6-A protocol,a series of samples that mixed by high and low level serum or urine pools with specific ratio were tested.To evaluate the anti-interference ability,we tested the sera added specific concentration of substances (bilirubin,hemoglobin,chyle).To verify the reasonable dilution ratio,diluted sera in different ratio were tested.Correlations and deviation among the detection results of six reagents were evaluated by testing 50 fresh patient samples.Accuracy was evaluated by measuring the NIST SRM 967a and NIST SRM 909b standard substances. Results There were no significant differences between the domestic and imported Cr reagents,with the within-run CV ranging from 0.5% to 1.2% and the total CV ranging from 0.5% to 1.9%,all of which were lower than the allowed biological variation for imprecision (3.0%).Linear ranges of the six Cr reagents were all relatively wide.No significant interference (bias%≤±10%) was found in the test of six reagents when conjugated bilirubin≤332μmol/L,unconjugated bilirubin≤327μmol/L,or chyle turbidity≤1530 FTU; no significant interference was found for the test of C,D,F reagents when Hb≤4.83 g/L; and for A,B,E when Hb≤3.381,3.864,4.347g/L,respectively.Percent deviation of the measured results of dulited serum samples diluted by 2 to 16 times compared to the results of the original samples ranged from 95.96% to 108.62%.Fresh patient samples comparison tests exhibited good correlation among the six reagents (r=1.000),although there were some differences (-4.91% to -4.75%).For NIST 967a,the percent bias ranged from -1.85% to 2.81% (level 1,66.5μmol/L),and from -2.28% to 2.05% (level 2,346.2μmol/L),none of the six reagents exceeded the allowed Cr bias derived from biological variation (4.0%).For NIST 909b,the percent bias ranged from -13.4% to 1.01% for level 1 (56.18μmol/L),while its value ranged from -6.49% to -1.79% for level 2 (467.4μmol/L),and these biases all exceeded the allowable range of target value. Conclusions Assessment of four domestic enzymatic Cr reagents and two imported reagents show high precision,wide linear range,and their dilution accuracy can meet the clinical needs.Moreover,the six regents show good anti-interference capability (bilirubin,hemoglobin,chyle) and high accuracy.No significant differences have been found in the above assessed properties among these reagents.(Chin J Lab Med,2013,36:806-811) Key words: Creatinine; Indicators and reagents; Reagent kits; diagnostic

Rapid detection of gene expression by a colorectal cancer Enzymatic Gene Chip Detection Kit

A weighted enzymatic chip array (WEnCA) can detect the gene expression of circulating tumor cells (CTCs) in the peripheral blood of patients with colorectal cancer (CRC). In this study, we used a reagent kit, the GeneCling CRC Enzymatic Gene Chip Detection Kit, which was specifically developed for the WEnCA operation platform to analyze the expression of 31 CRC-related genes of CTCs in the peripheral blood of 30 CRC patients. We moreover evaluated the expression of the genes by simultaneously using real-time quantitative polymerase chain reaction (RT-QPCR). The results showed the overexpression rate of nine genes—DVL1, ELAVL4, CHRNB1, UBE2C, PSAT1, CEA, PTTG1, KRT19, and hTERT—was greater than 90% in the CRC patients. This is concordant with the results of the original WEnCA operation procedure. Linear regression analysis demonstrated a high correlation (r = 0.901; p < 0.0001) between the experimental data of the detection kit and RT-QPCR. The GeneCling CRC Enzymatic Gene Chip Detection Kit is easy, fast, and convenient to operate for detecting gene expression of CTCs from peripheral blood. However, the correlation between the differential expression and the clinicopathological features of the 31 CRC-related genes needs further investigation to verify the clinical value.

Danthron activates AMP-activated protein kinase and regulates lipid and glucose metabolism in vitro

To discover the active compound on AMP-activated protein kinase (AMPK) activation and investigate the effects of the active compound 1,8-dihydroxyanthraquinone (danthron) from the traditional Chinese medicine rhubarb on AMPK-mediated lipid and glucose metabolism in vitro. HepG2 and C2C12 cells were used. Cell viability was determined using MTT assay. Real-time PCR was performed to measure the gene expression. Western blotting assay was applied to investigate the protein phosphorylation level. Enzymatic assay kits were used to detect the total cholesterol (TC), triglyceride (TG) and glucose contents. Danthron (0.1, 1, and 10 μmol/L) dose-dependently promoted the phosphorylation of AMPK and acetyl-CoA carboxylase (ACC) in both HepG2 and C2C12 cells. Meanwhile, danthron treatment significantly reduced the lipid synthesis related sterol regulatory element-binding protein 1c (SREBP1c) and fatty acid synthetase (FAS) gene expressions, and the TC and TG levels. In addition, danthron treatment efficiently increased glucose consumption. The actions of danthron on lipid and glucose metabolism were abolished or reversed by co-treatment with the AMPK inhibitor compound C. Danthron effectively reduces intracellular lipid contents and enhanced glucose consumption in vitro via activation of AMPK signaling pathway.

Abstract 285: The Effects of Hypercholesterolemia and Lipoprotein-Cholesterol Concentrations on Angiotensin II-Induced Abdominal Aortic Aneurysms

Background and objective Angiotensin II (AngII) infusion promotes abdominal aortic aneurysms (AAA) in both normo- and hypercholesterolemic mice, but the latter have a greater susceptibility, implicating a role of hypercholesterolemia in AngII-induced AAA. A recent publication demonstrated that high-density lipoprotein (HDL) administration prevented AngII-induced AAA in apoE -/- mice without affecting total plasma cholesterol concentrations. The purpose of this study was to determine the relationship between concentrations and characteristics of plasma cholesterol and lipoproteins and AngII-induced AAA. Methods and Results Three strains of male mice were studied: C57BL/6, LDL receptor -/-, and apoE -/-. These mice were fed either a normal diet or a saturated fat- enriched diet. AngII (1 μg/kg/min) was infused for 4 weeks through osmotic minipumps implanted subcutaneously. Total plasma cholesterol concentrations were measured using an enzymatic kit. Lipoproteins were resolved by size exclusion gel chromatography and non-HDL and HDL analyzed using Peakfit software. AAA was quantified by measuring maximal outer width of suprarenal aortas. In C57BL/6 mice, the fat-enriched diet modestly increased concentrations of total plasma cholesterol (114 ± 4 vs 99 ± 6 mg/dl; P=0.03) and HDL-C (98 ± 4 vs 81 ± 7 mg/dl; P=0.02), while did not change non-HDL-C/HDL-C ratio (0.21 ± 0.03 vs 0.24 ± 0.03; P=0.5) and AAA (1.1 ± 0.1 vs 1.1 ± 0.1 mm; P=0.6). In LDL receptor -/- mice, the fat-enriched diet feeding increased concentrations of total plasma cholesterol (1590 ± 58 vs 276 ± 9 mg/dl; P&lt; 0.001), HDL-C (132 ± 5 vs 90 ± 3 mg/dl; P&lt;0.001), non-HDL-C/HDL-C ratio (10.8 ± 0.3 vs 2.1 ± 0.1; P&lt;0.001), and AAA (1.6 ± 0.1 vs 1.2 ± 0.1 mm; P=0.002), compared to LDL receptor -/- mice fed the normal diet. In apoE -/- mice, the fat-enriched diet increased concentrations of total plasma cholesterol (1261 ± 121 vs 348 ± 38 mg/dl; P&lt;0.001), had no effect on HDL-C, increased non-HDL-C/HDL-C ratio (53.1 ± 5.9 vs 10.5 ± 2.0; P=0.016), and had no effect on AAA development (1.5 ± 0.2 vs 1.6 ± 0.2 mm; P=0.4). Conclusion Hypercholesterolemia promotes AngII-induced AAA, while AAA has no clear relationship with either concentrations of total plasma cholesterol or HDL-C.

BIOCHEMICAL AND SEROLOGICAL PROFILING OF SANDHI SHOOLA (ARTHRALGIA) PATIENTS OF AYURVEDA HOSPITAL

Sandhi shoola refers to the joints pain where there is pain with or without inflammation and can be caused by injury or a number of autoimmune disorders such as rheumatoid arthritis and gout. Ayurveda system of medicine believes this a s Sandhi shoola condition in which there is difficulty in walking and running and based on the physical examination , it is categorized into major types i.e. Aam v ata (AV) , S an dhi v ata (SV) and Vatar akta (V R) . However, biochemical and serologi cal investigations for Sandhi shoola were not cited in ancient Ayurveda literature . A retrospective study in which biochemical and serological investigations such as uric acid (UA), Ca 2+ and rheumatoid arthritis factor ( RAF), C - reactive p rotein (CRP), anti streptoly s in - O (ASO) were carried out respectively in 50 Sandhi shoola patients o f A yurveda hospital in Kolkata. The individual information about clinical symptoms and diagnosis by the hospital professionals were recorded. U A and Ca 2+ content in serum was determined by enzymatic and Cresolphthalein complexone kit methods respectiv ely, however RAF, CRP and ASO were determined thro ugh latex agglutination method. Results showed that maximum number of patients (24) suffered from AV followed by SV (15) and VR (1 1). The range of the UA and Ca 2+ were 2.6 – 18.0 mg/dl and 7.0 – 11.3 mg/dl respectively . T he average UA content is maximum in case of VR , while no significant differe nce was found in Ca 2+ content among the th ree Sandhi shoola . Serological tests indicated maximum RAF positive samples i n case of AV followed by S V p atients, while CRP was positive in maximum patients of AV and SV . Results revealed a strong diagnostic measure for Sandhi shoola and suggested that higher UA content and positive RAF should be a di agn ostic measure in case of Vatar akta and Aam v ata respectively. In conclusion, modern biochemical and serological investigations should be exercised in Ayurveda practice for diagnostic and validation with scientific justification strengthening the