Search results for: serum lipids
Commenced in January 2007
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Edition: International
Paper Count: 1181

Search results for: serum lipids

641 IL4/IL13 STAT6 Mediated Macrophage Polarization During Acute and Chronic Pancreatitis

Authors: Hager Elsheikh, Juliane Glaubitz, Frank Ulrich Weiss, Matthias Sendler

Abstract:

Aim: Acute pancreatitis (AP) and chronic pancreatitis (CP) are both accompanied by a prominent immune response which influences the course of disease. Whereas during AP the pro-inflammatory immune response dominates, during CP a fibroinflammatory response regulates organ remodeling. The transcription factor signal transducer and activator of transcription 6 (STAT6) is a crucial part of the Type 2 immune response. Here we investigate the role of STAT6 in a mouse model of AP and CP. Material and Methods: AP was induced by hourly repetitive i.p. injections of caerulein (50µg/kg/bodyweight) in C57Bl/6 J and STAT6-/- mice. CP was induced by repetitive caerulein injections 6 times a day, 3 days a week over 4 weeks. Disease severity was evaluated by serum amylase/lipase measurement, H&E staining of pancreas. Pancreatic infiltrate was characterized by immunofluorescent labeling of CD68, CD206, CCR2, CD4 and CD8. Pancreas fibrosis was evaluated by Azan blue staining. qRT-PCR was performed of Arg1, Nos2, Il6, Il1b, Col3a, Socs3 and Ym1. Affymetrix chip array analyses were done to illustrate the IL4/IL13/STAT6 signaling in bone marrow derived macrophages. Results: AP severity is mitigated in STAT6-/- mice, as shown by decreased serum amylase and lipase, as well as histological damage. CP mice surprisingly showed only slightly reduced fibrosis of the pancreas. Also staining of CD206 a classical marker of alternatively activated macrophages showed no decrease of M2-like polarization in the absence of STAT6. In contrast, transcription profile analysis in BMDM showed complete blockade of the IL4/IL13 pathway in STAT6-/- animals. Conclusion: STAT6 signaling pathway is protective during AP and mitigates the pancreatic damage. During chronic pancreatitis the IL4/IL13 – STAT6 axisis involved in organ fibrogenesis. Notably, fibrosis is not dependent on a single signaling pathway, and alternative macrophage activation is also complex and involves different subclasses (M2a, M2b, M2c and M2d) which could be independent of the IL4/IL13 STAT6 axis.

Keywords: chronic pancreatitis, macrophages, IL4/IL13, Type immune response

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640 Utility of Cardiac Biomarkers in Combination with Exercise Stress Testing in Patients with Suspected Ischemic Heart Disease

Authors: Rawa Delshada, Sanaa G. Hamab, Rastee D. Koyeec

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Eighty patients with suspected ischemic heart disease were enrolled in the present study. They were classified into two groups: patients with positive exercise stress test results (n=40) and control group with negative exercise stress test results (n=40). Serum concentration of troponin I, Heart-type Fatty Acid Binding Protein (H-FABP) and Ischemia Modified Albumin (IMA) were measured one hour after performing stress test. Enzyme Linked Immunosorbent Assay was used to measure both troponin I, H-FABP levels, while IMA levels were measured by albumin cobalt binding test. There was no statistically significant difference in the mean concentration of troponin I between two groups (0.75±0.55ng/ml) for patients with positive test result vs. (0.71±0.55ng/ml) for negative test result group with P>0.05. Contrary to our expectation, mean IMA level was slightly higher among control group (70.88±39.76U/ml) compared to (62.7±51.9U/ml) in positive test result group, but still with no statistically significant difference (P>0.05). Median H-FABP level was also higher among negative exercise stress testing group compared the positive one (2ng/ml vs. 1.9ng/ml respectively), but failed to reach statistically significant difference (P>0.05). When quartiles model used to explore the possible association between each study biomarkers with the others; serum H-FABP level was lowest (1.7ng/ml) in highest quartile of IMA and lowest H-FABP (1.8ng/ml) in highest quartile of troponin I but with no statistically significant association (P>0.05). Myocardial ischemia, more likely occurred after exercise stress test, is not capable of causing troponin I release. Furthermore, an increase in H-FABP and IMA levels after stress test are not reflecting myocardial ischemia. Moreover, the combination of troponin I, H-FABP and IMA after measuring their post exercise levels does not improve the diagnostic utility of exercise stress test enormously.

Keywords: cardiac biomarkers, ischemic heart disease, troponin I, ischemia modified albumin, heart-type fatty acid binding protein, exercise stress testing

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639 Sialic Acid Profile and Sialidase Activity in HIV-Infected Individuals

Authors: Hadiza Abdullahi

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Sialic Acids and sialidases have been implicated in many disease states particularly bacterial and viral infections which are common opportunist infections of HIV disease. Their role in HIV/AIDS is contemplated. A study was carried out to determine Sialic Acid profile and Sialidase Activity in HIV infected and Apparently Healthy individuals, and also determine the relationship between the sialic acid levels and sialidase activity. Blood samples were collected from 200 subjects (150 HIV infected individuals and 50 apparently healthy individuals divided into four groups- HIV ART Naïve, HIV Stable (on ART but have been stable with no clinical episodes), HIV-OI (on ART with opportunistic infections), and Apparently Healthy). Complete Blood Count, Erythrocyte Surface Sialic Acid (ESSA), Free Serum Sialic Acid (FSSA) concentrations and Sialidase activity were determined for all 200 subjects. Analysis of variance (ANOVA) was used to compare the results of the different groups of HIV infected individuals as well as controls. The mean haemoglobin (HGB), Packed Cell Volume (PCV) and Red Blood Cells (RBC) concentrations were significantly lower (P ≤ 0.05) in the HIV groups compared with the apparently healthy groups. Anaemia and neutropaenia were the most common heamatological abnormalities observed in this study with highest prevalence of anaemia found in the ART naive group. The mean FSSA was 0.4±0.4mg/ml. There was a significant difference (p ≤ 0.05) between some groups. The highest levels of FSSA was observed in the HIV ART naïve (0.65±0.5mg/ml). The mean ESSA value for the study population was 0.54±0.35mg/ml with no significant difference (p ≤ 0.05) between groups. The mean sialidase activity values were 0.52±0.1 µmol/min/µl, 0.40±0.1 µmol/min/µl, 0.45±0.1 µmol/min/µl and 0.41±0.1 µmol/min/µl for the HIV ART naïve, HIV stable, HIV/OIs and apparently healthy groups respectively. No significant difference (p ≤ 0.05) was found between groups and also in gender and age. The finding in this study of higher mean sialidase activity and FSSA levels in the ART naïve HIV group compared with other groups indicate that the virus and other opportunistic pathogens may be sialidase producers in vivo which cleave off sialic acids from erythrocytes surface, leading to high levels of FSSA, anaemia and neutropaenia seen in this group. The higher ESSA concentration found in the HIV stable group along with lowest FSSA concentration in the group suggests the presence of sialyltransferases.

Keywords: erythrocyte surface sialic acid, free serum sialic acid, HIV, sialidase

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638 Delivery of Doxorubicin to Glioblastoma Multiforme Using Solid Lipid Nanoparticles with Surface Aprotinin and Melanotransferrin Antibody for Enhanced Chemotherapy

Authors: Yung-Chih Kuo, I-Hsuan Lee

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Solid lipid nanoparticles (SLNs) conjugated with aprotinin (Apr) and melanotransferrin antibody (Anti-MTf) were used to carry doxorubicin (Dox) across the blood–brain barrier (BBB) for glioblastoma multiforme (GBM) chemotherapy. Dox-entrapped SLNs with grafted Apr and Anti-MTf (Apr-Anti-MTf-Dox-SLNs) were applied to a cultured monolayer comprising human brain-microvascular endothelial cells (HBMECs) with regulation of human astrocyte (HAs) and to a proliferated colony of U87MG cells. Based on the average particle diameter, zeta potential, entrapping efficiency of Dox, and grafting efficiency of Apr and Anti-MTf, we found that 40% (w/w) 1,2-dipalmitoyl-sn-glycero-3-phosphocholine in lipids were appropriate for fabricating Apr-Anti-MTf-Dox-SLNs. In addition, Apr-Anti-MTf-Dox-SLNs could prevent Dox from fast dissolution and did not induce a serious cytotoxicity to HBMECs and HAs when compared with free Dox. Moreover, the treatments with Apr-Anti-MTf-Dox-SLNs enhanced the ability of Dox to infuse the BBB and to inhibit the growth of GBM. The current Apr-Anti-MTf-Dox-SLNs can be a promising pharmacotherapeutic preparation to penetrate the BBB for malignant brain tumor treatment.

Keywords: solid lipid nanoparticle, glioblastoma multiforme, blood–brain barrier, doxorubicin

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637 Evaluation of Antibody Titer Produced in Layer Chicken after Vaccination with an Experimental Ornitobacterium rhinotracheal Vaccine

Authors: Mohammad Javad Mehrabanpour, Mohammad Hosein Hosseini, Ali Shirazi, Dorsa Mehrabanpour

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Respiratory infections are the most important diseases that affect poultry. Ornithobacterium rhinotracheale is a bacterium that causes respiratory infections including alveolar inflation and pneumonia in birds. The aim of this study was to evaluated antibody titer against Ornitobacterium rhinotracheal in layer chicken sera after vaccination with an experimental ORT vaccine that produced in Razi Vaccine and Serum Research Institute. Cultured bacteria were inactivated by formalin, and controlled tests were conducted on it. The obtained antigens were formulated using Montanide oil and were homogenized using homogenizer. Eighty SPF chickens were kept until the age of 14 days under existing standards for temperature, humidity, and light. At the age of 14 days, chickens were divided into 3 groups. The first group included 50 chickens injected with prepared ORT vaccine, the second group, as control group, included 15 chickens injected with sterile PBS to get stress of infection and the third group included 15 chickens with no injection performed to them. All 3 groups were kept in separate cages at same room. Blood samples were regularly taken from the chickens every week for serum separation and evaluation of antibody titer. During the fifth week post vaccination, booster vaccine was injected into the chickens of vaccinated group. The chickens were inspected every day in terms of mortality as well as any injection site reactions. Three weeks after the booster injection, blood samples were taken from all chickens of all groups, and sera were isolated. The sera of immunized (vaccinated) SPF chickens with ORT vaccine as well as that of SPF chickens in the control groups were reviewed according to the recommendations of ELISA kit manufacturer to examine the chicken’s humeral immune response to the studied vaccine. Potency, stability and sterility tests were also performed on the above mentioned vaccine. Results obtained indicate high antibody titer in sera of chickens vaccinated with experimental ORT vaccine as compared with the control groups that emphasize the ability of experimentally prepared ORT vaccine to stimulate humoral immune response of chicken. After the second injection, antibody titer increased and remained almost stable up to 9 weeks after the injection. ORT vaccine can cause potency in chickens and can protect them against disease.

Keywords: antibody, layer chicken, Ornithobactrium rhinotracitis, vaccine

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636 Increased Risk of Adverse Birth Outcomes of Newborns in Arsenic Exposed- Women with Gestational Diabetes

Authors: Tania Mannan, Rahelee Zinnat, Fatema Jebunnesa, Israt Ara Hossain

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Background: Exposure to arsenic has known toxic effects but the effect on pregnancy outcomes is not as widely documented especially in women with diabetes. Growing evidence has suggested a potential role of arsenic exposure in the development of gestational diabetes mellitus (GDM). Therefore, we aimed to investigate the association of urinary arsenic (UAs) with birth outcomes in GDM subjects. Methods: Under an observational cross-sectional design a total of 263 GDM subjects (age in years, M±SD, 21±3.7) residing in an arsenic affected area of Bangladesh, were subjected to a 2 sample OGTT at the third trimester of gestation. Among them, 73 GDM and 190 non-GDM subjects enrolled in this study. Clinical and anthropometric measurements were done by standard techniques. Degree of chronic arsenic exposure was assessed by the level of UAs level. According to World Health Organization (WHO) criteria, GDM was diagnosed and neonatal outcomes using APGAR (Activity Pulse Grimace Appearance Respirations) Score, birth weight and size were assessed by a specialist obstetrician. Serum glucose was measured by the Glucose Oxidase method and UAs level was determined by ultraviolet/visible spectrophotometry. Result: Out of the 263 pregnant women, 28% developed GDM. Urinary Arsenic was significantly higher in the GDM as compared to the non-GDM group [UAs, µg/l, M±SD (range), 204.2±67.0 (67.0-377.0) vs 77.3±38.1 (22.0-99.0), p < 0.001]. Activity Pulse Grimace Appearance Respirations Score of the neonates from GDM mothers was significantly lower compared to the neonates from non-GDM mothers [APGAR Score, M±SD, 4.7±0.8 vs. 6.4±0.7, p<0.001]. Pearson’s correlation analysis in GDM subjects revealed that UA levels were found to have a significant positive correlation with both fasting and postprandial serum glucose levels (p < 0.001) and (p < 0.001) respectively. Again, a significant inverse correlation of UAs with birth weight and size was observed (p < 0.001). The APGAR Score of the neonates were found to have a significant negative correlation (p < 0.001) with UAs level. Conclusion: The effect of chronic arsenic exposure is associated with glucose intolerance during pregnancy and it also adversely affects birth outcomes. The study suggests further research on the impact of total arsenic exposure on pregnancy outcomes.

Keywords: APGAR score, arsenic exposure, birth outcome, gestational diabetes mellitus,

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635 Microbial Removal of Polycyclic Aromatic Hydrocarbons from Petroleum Refinery Sludge: A Consortial Approach

Authors: Dheepshika Kodieswaran

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The persisting problem in the world that continuously impose our planet at risk is the increasing amounts of recalcitrant. One such issue is the disposal of the Petroleum Refinery Sludge (PRS) which constitutes hydrocarbons that are hazardous to terrestrial and aquatic life. The comparatively safe approach to handling these wastes is by microbial degradation, while the other chemical and physical methods are either expensive and/or produce secondary pollutants. The bacterial and algal systems have different pathways for the degradation of hydrocarbons, and their growth rates vary. This study shows how different bacterial and microalgal strains degrade the polyaromatic hydrocarbon PAHs individually and their symbiotic influence on degradation as well. In this system, the metabolites and gaseous exchange help each other in growth. This method using also aids in the accumulation of lipids in microalgal cells and from which bio-oils can also be extracted. The bacterial strains used in this experiment are reported to be indigenous strains isolated from PRS. The target PAH studied were anthracene and pyrene for a period of 28 days. The PAH degradation kinetics best fitted the Gompertz model, and the order of the kinetics, rate constants, and half-life was determined.

Keywords: petroleum refinery sludge, co-culturing, polycyclic hydrocarbons, microalgal-bacterial consortia

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634 Glyco-Biosensing as a Novel Tool for Prostate Cancer Early-Stage Diagnosis

Authors: Pavel Damborsky, Martina Zamorova, Jaroslav Katrlik

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Prostate cancer is annually the most common newly diagnosed cancer among men. An extensive number of evidence suggests that traditional serum Prostate-specific antigen (PSA) assay still suffers from a lack of sufficient specificity and sensitivity resulting in vast over-diagnosis and overtreatment. Thus, the early-stage detection of prostate cancer (PCa) plays undisputedly a critical role for successful treatment and improved quality of life. Over the last decade, particular altered glycans have been described that are associated with a range of chronic diseases, including cancer and inflammation. These glycans differences enable a distinction to be made between physiological and pathological state and suggest a valuable biosensing tool for diagnosis and follow-up purposes. Aberrant glycosylation is one of the major characteristics of disease progression. Consequently, the aim of this study was to develop a more reliable tool for early-stage PCa diagnosis employing lectins as glyco-recognition elements. Biosensor and biochip technology putting to use lectin-based glyco-profiling is one of the most promising strategies aimed at providing fast and efficient analysis of glycoproteins. The proof-of-concept experiments based on sandwich assay employing anti-PSA antibody and an aptamer as a capture molecules followed by lectin glycoprofiling were performed. We present a lectin-based biosensing assay for glycoprofiling of serum biomarker PSA using different biosensor and biochip platforms such as label-free surface plasmon resonance (SPR) and microarray with fluorescent label. The results suggest significant differences in interaction of particular lectins with PSA. The antibody-based assay is frequently associated with the sensitivity, reproducibility, and cross-reactivity issues. Aptamers provide remarkable advantages over antibodies due to the nucleic acid origin, stability and no glycosylation. All these data are further step for construction of highly selective, sensitive and reliable sensors for early-stage diagnosis. The experimental set-up also holds promise for the development of comparable assays with other glycosylated disease biomarkers.

Keywords: biomarker, glycosylation, lectin, prostate cancer

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633 Oil and Proteins of Sardine (Sardina Pilchardus) Compared with Casein or Mixture of Vegetable Oils Improves Dyslipidemia and Reduces Inflammation and Oxidative Stress in Hypercholesterolemic and Obese Rats

Authors: Khelladi Hadj Mostefa, Krouf Djamil, Taleb-Dida Nawel

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Background: Obesity results from a prolonged imbalance between energy intake and energy expenditure, as depending on basal metabolic rate. Oils and proteins from sea have important therapeutic (such as obesity and hypercholesterolemia) and antioxidant effects. Sardine are a widely consumed fish in the Mediterranean region. Its consumption provides humans with various nutrients such as oils (rich in omega 3 plyunsaturated fatty acids)) and proteins. Methods: Sardine oil (SO) and sardine proteins (SP) were extracted and purified. Mixture of vegetable oils (olive-walnut-sunflower) were prepared from oils produced in Algeria. Eighteen wistar rats are fed a high fat diet enriched with 1% cholesterol for 30 days to induce obesity and hypercholesterolemia. The rats are divided into 3 groups. The first group consumes 20% sardine protein combined with 5% sardine oil (38% SFA (saturated fatty acids), 31% MIFA (monounsaturated fatty acids) and 31% PIFA (polyunsaturated fatty acids)) (SPso). The second group consumes 20% sardine protein combined with 5% of a mixture of vegetable oils (VO) containing 13% SFA, 58% MIFA and 29% PIFA (PSvo), and the third group consuming 20% casein combined with 5% of the mixture of vegetable oils and serves as a semi-synthetic reference (CASvo). Body weights and glycaemia are measured weekly After 28 days of experimentation, the rats are sacrificed, the blood and the liver removed. Serum assays of total cholesterol (TC) and triglycerides (TG) were performed by enzymatic colorimetric methods. Evaluation of lipid peroxidation was performed by assaying thiobarbituric acid reactive species (TBARS) and hydroperoxides values. The protein oxidation was performed by assaying carbonyl derivatives values. Finally, evaluation of antioxidant defense is made by measuring the activity of antioxidant enzymes, the superoxide dismutase (SOD) and the catalase (CAT).Results: After 28 days, the body weight (BW) of the rats increased significantly in SPso and SPvo groups compared to CAS group, by +11% and 7%, respectively. Cholesterolemia (TC) increased significantly in the SPso and SPvo groups compared to the CAS group (P<0.01), while triglyceridemia (TG) decreased significantly in the SPso group compared to SPvo and CAS groups (P<0.01). Albumin (marker of inflammation) increased in the PSs group compared to SPvo and CAS groups by +35% and +13%, respectively. The serum TBARS levels are -40% lower in SPso group compared to SPvo group, and they are -80% and -76% lower in SPso compared to SPvo and CAS groups, respectively. The level of carbonyls derivatives in the serum and liver are significantly reduced in the SPso group compared to the SPvo and CAS groups. Superoxide dismutase (SOD) activity decreased in liver of SPso group compared to SPvo group (P<0.01). While that of CAT is increased in liver tissue of SPso group compared to SPvo group (P<0.01). Conclusion: Sardine oil combined with sardine protein has a hypotriglyceridemic effect, reduces body weight, attenuates inflammation and seems to protect against lipid peroxidation and protein oxidation and increases antioxidant defense in hypercholesterolemic and obese rats. This could be in favor of a protective effect against obesity and cardiovascular diseases.

Keywords: rat, obesity, hypercholesterolemia, sardine protein, sardine oil, vegetable oils mixture, lipid peroxidation, protein oxidation, antioxidant defense

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632 Hypocalcaemia Inducing Heart Failure: A Rare Presentation

Authors: A. Kherraf, M. Bouziane, L. Azzouzi, R. Habbal

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Introduction: Hypocalcaemia is a rare cause of heart failure. We report the clinical case of a young patient with reversible dilated cardiomyopathy secondary to hypocalcaemia in the context of hyperparathyroidism. Clinical case: We report the clinical case of a 23-year-old patient with a history of thyroidectomy for papillary thyroid carcinoma 3 years previously, who presented to the emergency room with a progressive onset dyspnea and edema of the lower limbs. Clinical examination showed hypotension at 90/70 mmHg, tachycardia at 102 bpm, and edema of the lower limbs. The ECG showed a regular sinus rhythm with a prolonged corrected QT interval to 520ms. The chest x-ray showed cardiomegaly. Echocardiography revealed dilated cardiomyopathy with biventricular dysfunction and a left ventricular ejection fraction of 45%, as well as moderate mitral insufficiency by restriction of the posterior mitral leaflet, moderate tricuspid insufficiency, and a dilated inferior vena cava with a pulmonary arterial pressure estimated at 46 mmHg. Blood tests revealed severe hypocalcemia at 38 mg / l with normal albumin and thyroxine levels, as well as hyperphosphatemia and increased TSH. The patient received calcium intake and vitamin D supplementation and was treated with beta blockers, ACE inhibitors, and diuretics with good progress and progressive normalization of cardiac function. Discussion: The cardiovascular manifestations of hypocalcaemia usually appear with deeply low serum calcium levels. This can lead to hypotension, arrhythmias, ventricular fibrillation, prolonged QT interval, or even heart failure. Heart failure is a rare and serious complication of hypocalcemia but most often characterized by complete normalization of myocardial function after treatment. The etiology of the hypocalcaemia, in this case, was probably related to accidental parathyroid removal during thyroidectomy. This is why careful monitoring of calcium levels is recommended after surgery. Conclusion: Hypocalcemic heart failure is rare but reversible heart disease. Systematic monitoring of serum calcium should be performed in all patients after thyroid surgery to avoid any complications related to hypoparathyroidism.

Keywords: hypocalcemia, heart failure, thyroid surgery, hypoparathyroidism

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631 Formulation and Evaluation of Curcumin-Zn (II) Microparticulate Drug Delivery System for Antimalarial Activity

Authors: M. R. Aher, R. B. Laware, G. S. Asane, B. S. Kuchekar

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Objective: Studies have shown that a new combination therapy with Artemisinin derivatives and curcumin is unique, with potential advantages over known ACTs. In present study an attempt was made to prepare microparticulate drug delivery system of Curcumin-Zn complex and evaluate it in combination with artemether for antimalarial activity. Material and method: Curcumin Zn complex was prepared and encapsulated using sodium alginate. Microparticles thus obtained are further coated with various enteric polymers at different coating thickness to control the release. Microparticles are evaluated for encapsulation efficiency, drug loading and in vitro drug release. Roentgenographic Studies was conducted in rabbits with BaSO 4 tagged formulation. Optimized formulation was screened for antimalarial activity using P. berghei-infected mice survival test and % paracetemia inhibition, alone (three oral dose of 5mg/day) and in combination with arthemether (i.p. 500, 1000 and 1500µg). Curcumin-Zn(II) was estimated in serum after oral administration to rats by using spectroflurometry. Result: Microparticles coated with Cellulose acetate phthalate showed most satisfactory and controlled release with 479 min time for 60% drug release. X-ray images taken at different time intervals confirmed the retention of formulation in GI tract. Estimation of curcumin in serum by spectroflurometry showed that drug concentration is maintained in the blood for longer time with tmax of 6 hours. The survival time (40 days post treatment) of mice infected with P. berghei was compared to survival after treatment with either Curcumin-Zn(II) microparticles artemether combination, curcumin-Zn complex and artemether. Oral administration of Curcumin-Zn(II)-artemether prolonged the survival of P.berghei-infected mice. All the mice treated with Curcumin-Zn(II) microparticles (5mg/day) artemether (1000µg) survived for more than 40 days and recovered with no detectable parasitemia. Administration of Curcumin-Zn(II) artemether combination reduced the parasitemia in mice by more than 90% compared to that in control mice for the first 3 days after treatment. Conclusion: Antimalarial activity of the curcumin Zn-artemether combination was more pronounced than mono therapy. A single dose of 1000µg of artemether in curcumin-Zn combination gives complete protection in P. berghei-infected mice. This may reduce the chances of drug resistance in malaria management.

Keywords: formulation, microparticulate drug delivery, antimalarial, pharmaceutics

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630 Therapeutic Effect of Indane 1,3-Dione Derivatives in the Restoration of Insulin Resistance in Human Liver Cells and in Db/Db Mice Model: Biochemical, Physiological and Molecular Insights of Investigation

Authors: Gulnaz Khan, Meha F. Aftab, Munazza Murtaza, Rizwana S. Waraich

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Advanced glycation end products (AGEs) precursor and its abnormal accumulation cause damage to various tissues and organs. AGEs have pathogenic implication in several diseases including diabetes. Existing AGEs inhibitors are not in clinical use, and there is a need for development of novel inhibitors. The present investigation aimed at identifying the novel AGEs inhibitors and assessing their mechanism of action for treating insulin resistance in mice model of diabetes. Novel derivatives of benzylidene of indan-1,3-dione were synthesized. The compounds were selected to study their action mechanism in improving insulin resistance, in vitro, in human hepatocytes and murine adipocytes and then, in vivo, in mice genetic model of diabetes (db/db). Mice were treated with novel derivatives of benzylidene of indane 1,3-dione. AGEs mediated ROS production was measured by dihydroethidium fluorescence assay. AGEs level in the serum of treated mice was observed by ELISA. Gene expression of receptor for AGEs (RAGE), PPAR-gamma, TNF-alpha and GLUT-4 was evaluated by RT-PCR. Glucose uptake was measured by fluorescent method. Microscopy was used to analyze glycogen synthesis in muscle. Among several derivatives of benzylidene of indan-1,3-dione, IDD-24, demonstrated highest inhibition of AGESs. IDD-24 significantly reduced AGEs formation and expression of receptor for advanced glycation end products (RAGE) in fat, liver of db/db mice. Suppression of AGEs mediated ROS production was also observed in hepatocytes and fat cell, after treatment with IDD-24. Glycogen synthesis was increased in muscle tissue of mice treated with IDD-24. In adipocytes, IDD-24 prevented AGEs induced reduced glucose uptake. Mice treated with IDD-24 exhibited increased glucose tolerance, serum adiponectin levels and decreased insulin resistance. The result of present study suggested that IDD-24 can be a possible treatment target to address glycotoxins induced insulin resistance.

Keywords: advance glycation end product, hyperglycemia, indan-1, 3-dione, insulin resistance

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629 Study of the Biological Activity of a Ganglioside-Containing Drug (Cronassil) in an Experimental Model of Multiple Sclerosis

Authors: Hasmik V. Zanginyan, Gayane S. Ghazaryan, Laura M. Hovsepyan

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Experimental autoimmune encephalomyelitis (EAE) is an inflammatory demyelinating disease of the central nervous system that is induced in laboratory animals by developing an immune response against myelin epitopes. The typical clinical course is ascending palsy, which correlates with inflammation and tissue damage in the thoracolumbar spinal cord, although the optic nerves and brain (especially the subpial white matter and brainstem) are also often affected. With multiple sclerosis, there is a violation of lipid metabolism in myelin. When membrane lipids (glycosphingolipids, phospholipids) are disturbed, metabolites not only play a structural role in membranes but are also sources of secondary mediators that transmit multiple cellular signals. The purpose of this study was to investigate the effect of ganglioside as a therapeutic agent in experimental multiple sclerosis. The biological activity of a ganglioside-containing medicinal preparation (Cronassial) was evaluated in an experimental model of multiple sclerosis in laboratory animals. An experimental model of multiple sclerosis in rats was obtained by immunization with myelin basic protein (MBP), as well as homogenization of the spinal cord or brain. EAE was induced by administering a mixture of an encephalitogenic mixture (EGM) with Complete Freund’s Adjuvant. Mitochondrial fraction was isolated in a medium containing 0,25 M saccharose and 0, 01 M tris buffer, pH - 7,4, by a method of differential centrifugation on a K-24 centrifuge. Glutathione peroxidase activity was assessed by reduction reactions of hydrogen peroxide (H₂O₂) and lipid hydroperoxides (ROOH) in the presence of GSH. LPO activity was assessed by the amount of malondialdehyde (MDA) in the total homogenate and mitochondrial fraction of the spinal cord and brain of control and experimental autoimmune encephalomyelitis rats. MDA was assessed by a reaction with Thiobarbituric acid. For statistical data analysis on PNP, SPSS (Statistical Package for Social Science) package was used. The nature of the distribution of the obtained data was determined by the Kolmogorov-Smirnov criterion. The comparative analysis was performed using a nonparametric Mann-Whitney test. The differences were statistically significant when р ≤ 0,05 or р ≤ 0,01. Correlational analysis was conducted using a nonparametric Spearman test. In the work, refrigeratory centrifuge, spectrophotometer LKB Biochrom ULTROSPECII (Sweden), pH-meter PL-600 mrc (Israel), guanosine, and ATP (Sigma). The study of the process of lipid peroxidation in the total homogenate of the brain and spinal cord in experimental animals revealed an increase in the content of malonic dialdehyde. When applied, Cronassial observed normalization of lipid peroxidation processes. Reactive oxygen species, causing lipid peroxidation processes, can be toxic both for neurons and for oligodendrocytes that form myelin, causing a violation of their lipid composition. The high content of lipids in the brain and the uniqueness of their structure determines the nature of the development of LPO processes. The lipid layer of cellular and intracellular membranes performs two main functions -barrier and matrix (structural). Damage to the barrier leads to dysregulation of intracellular processes and severe disorders of cellular functions.

Keywords: experimental autoimmune encephalomyelitis, multiple sclerosis, neuroinflammation, therapy

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628 Novel p22-Monoclonal Antibody Based Blocking ELISA for the Detection of African Swine Fever Virus Antibodies in Serum

Authors: Ghebremedhin Tsegay, Weldu Tesfagaber, Yuanmao Zhu, Xijun He, Wan Wang, Zhenjiang Zhang, Encheng Sun, Jinya Zhang, Yuntao Guan, Fang Li, Renqiang Liu, Zhigao Bu, Dongming Zhao*

Abstract:

African swine fever (ASF) is a highly infectious viral disease of pigs, resulting in significant economic loss worldwide. As there is no approved vaccines and treatments, the control of ASF entirely depends on early diagnosis and culling of infected pigs. Thus, highly specific and sensitive diagnostic assays are required for accurate and early diagnosis of ASF virus (ASFV). Currently, only a few recombinant proteins have been tested and validated for use as reagents in ASF diagnostic assays. The most promising ones for ASFV antibody detection were p72, p30, p54, and pp62. So far, three ELISA kits based on these recombinant proteins have been commercialized. Due to the complex nature of the virus and variety forms of the disease, robust serodiagnostic assays are still required. ASFV p22 protein, encoded by KP177R gene, is located in the inner membrane of viral particle and appeared transiently in the plasma membrane early after virus infection. The p22 protein interacts with numerous cellular proteins, involved in processes of phagocytosis and endocytosis through different cellular pathways. However, p22 does not seem to be involved in virus replication or swine pathogenicity. In this study, E.coli expressed recombinant p22 protein was used to generate a monoclonal antibody (mAb), and its potential use for the development of blocking ELISA (bELISA) was evaluated. A total of 806 pig serum samples were tested to evaluate the bELISA. Acording the ROC (Reciever operating chracteristic) analysis, 100% sensitivity and 98.10% of specificity was recorded when the PI cut-off value was set at 47%. The novel assay was able to detect the antibodies as early as 9 days post infection. Finaly, a highly sensitive, specific and rapid novel p22-mAb based bELISA assay was developed, and optimized for detection of antibodies against genotype I and II ASFVs. It is a promising candidate for an early and acurate detection of the antibodies and is highly expected to have a valuable role in the containment and prevention of ASF.

Keywords: ASFV, blocking ELISA, diagnosis, monoclonal antibodies, sensitivity, specificity

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627 Isothermal Crystallization Kinetics of Lauric Acid Methyl Ester from DSC Measurements

Authors: Charine Faith H. Lagrimas, Rommel N. Galvan, Rizalinda L. de Leon

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An ongoing study, methyl laurate to be used as a refrigerant in an HVAC system, requires the crystallization kinetics of the said substance. Step-wise and normal forms of Avrami model parameters were used to describe the isothermal crystallization kinetics of methyl laurate at different temperatures from Differential Scanning Calorimetry (DSC) measurements. At 3 °C, parameters showed that methyl laurate exhibits a secondary crystallization. The primary crystallization occurred with instantaneous nuclei and spherulitic growth; followed by a secondary instantaneous nucleation with a lower growth of dimensionality, rod-like. At 4 °C to 6 °C, the exotherms from DSC implied that the system was under the isokinetic range. The kinetics behavior is the same which is instantaneous nucleation with one-dimensional growth. The differences for the isokinetic range temperatures are the activation energies (directly proportional to T) and nucleation rates (inversely proportional to T). From the images obtained during the crystallization of methyl laurate using an optical microscope, it is confirmed that the nucleation and crystal growth modes obtained from the optical microscope are consistent with the parameters from Avrami model.

Keywords: Avrami model, isothermal crystallization, lipids kinetics, methyl laurate

Procedia PDF Downloads 331
626 Comparison Serum Vitamin D by Geographic between the Highland and Lowland Schizophrenic Patient in the Sumatera Utara

Authors: Novita Linda Akbar, Elmeida Effendy, Mustafa M. Amin

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Background: The most common of psychotic disorders is schizophrenia. Vitamin D is made from sunlight, and in the skin from UVB radiation from sunlight. If people with Vitamin D deficiency is common severe mental illness such as schizophrenia.Schizophrenia is a chronic mental illness characterised by positive symptoms and negatives symptoms, such as hallucinations and delusions, flat affect and lack of motivation we can found. In patients with Schizophrenia maybe have several environmental risk factors for schizophrenia, such as season of birth, latitude, and climate has been linked to vitamin D deficiency. There is also relationship between the risk of schizophrenia and latitude, and with an increased incidence rate of schizophrenia seen at a higher latitude. Methods: This study was an analytical study, conducted in BLUD RS Jiwa Propinsi Sumatera Utara and RSUD Deli Serdang, the period in May 2016 and ended in June 2016 with a sample of the study 60 sample (20 patients live in the Highland and Lowland, 20 healthy controls). Inclusion criteria were schizophrenic patients both men and women, aged between 18 to 60 years old, acute phase no agitation or abstinence antipsychotic drugs for two weeks, live in the Highland and Lowland, and willing to participate this study. Exclusion criteria were history of other psychotic disorders, comorbidities with other common medical condition, a history of substance abuse. Sample inspection for serum vitamin D using ELFA method. Statistical analysis using numeric comparative T-independent test. Results: The results showed that average levels of vitamin D for a group of subjects living in areas of high land was 227.6 ng / mL with a standard deviation of 86.78 ng / mL, the lowest levels of vitamin D is 138 ng / mL and the highest 482 ng / mL. In the group of subjects who settled in the low lands seem mean vitamin D levels higher than the mountainous area with an average 237.8 ng / mL with a standard deviation of 100.16 ng / mL. Vitamin D levels are lowest and the highest 138-585 ng / mL. Conclusion and Suggestion: The results of the analysis using the Mann Whitney test showed that there were no significant differences between the mean for the levels of vitamin D based on residence subject with a value of p = 0.652.

Keywords: latitude, schizophrenia, Vitamin D, Sumatera Utara

Procedia PDF Downloads 247
625 Microbial and Meiofaunal Dynamics in the Intertidal Sediments of the Northern Red Sea

Authors: Hamed A. El-Serehy, Khaled A. Al-Rasheid, Fahad A Al-Misned

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The meiofaunal population fluctuation, microbial dynamic and the composition of the sedimentary organic matter were investigated seasonally in the Egyptian shores along the northern part of Red Sea. Total meiofaunal population densities were extremely low with an annual average of 109 ±26 ind./10 cm2 and largely dominated by nematodes (on annual average from 52% to 94% of total meiofaunal density). The benthic microbial population densities ranged from 0.26±0.02 x 108 to 102.67±18.62 x 108/g dry sediment. Total sedimentary organic matter concentrations varied between 5.8 and 11.6 mg/g and the organic carbon, which was measured as summation of the carbohydrates, proteins and lipids, accounted for only a small fraction of being 32 % of the total organic matter. Chlorophyll a attained very low values and fluctuated between 2 and 11 µg/g. The very low chlorophyll a concentration in the Egyptian coasts along the Red Sea can suggest that the sedimentary organic matter along the Egyptian coasts is dominated by organic detrital and heterotrophic bacteria on one hand, and do not promote carbon transfer towards the higher trophic level on the other hand. However, the present study indicates that the existing of well diversified meiofaunal group, with a total of ten meiofaunal taxa, can serve as food for higher trophic levels in the Red Sea marine ecosystem.

Keywords: bacteria, meiofauna, intertidal sediments, Red Sea

Procedia PDF Downloads 416
624 Application of Coaxial Electrospinning for the Encapsulation of Omega-3 Fatty Acids

Authors: Sokratis Koskinakis, Georgia Frakolaki, Magdalini Krokida

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Regular consumption of omega-3 fatty acid-rich lipids is said to provide a wide range of health benefits, including prevention of inflammation, cardiovascular disease, diabetes, arthritis, and ulcerative colitis. Because of their potential nutritional and health benefits, the omega-3 PUFAs are increasingly being supplemented in functional food products meant to improve human health and wellbeing. However, dietary fortification with PUFAs is difficult due to their low water solubility, tendency to oxidize quickly, and inconsistent bioavailability. These issues can be solved through application of modern encapsulation technologies, which typically entail integrating omega-3 oils into well-designed matrices made from food-grade components. Electrospinning, for example, is an effective encapsulation method for producing sub-micron or nano-scale polymer fibers. For this purpose, various combinations of hydroxypropyl-β-cyclodextrin and cellulose nanocrystals/ nanofibers were assessed for the encapsulation of omega-3 fatty acids through the innovative technology of electrospinning. The encapsulation yield was evaluated through GC-analysis, and the morphology of the final products was assessed through SEM analysis.

Keywords: electrospinning, encapsulation, omega-3 fatty acids, cellulose nanocrystals / nanofibers

Procedia PDF Downloads 70
623 A Dynamic Model for Assessing the Advanced Glycation End Product Formation in Diabetes

Authors: Victor Arokia Doss, Kuberapandian Dharaniyambigai, K. Julia Rose Mary

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Advanced Glycation End (AGE) products are the end products due to the reaction between excess reducing sugar present in diabetes and free amino group in protein lipids and nucleic acids. Thus, non-enzymic glycation of molecules such as hemoglobin, collagen, and other structurally and functionally important proteins add to the pathogenic complications such as diabetic retinopathy, neuropathy, nephropathy, vascular changes, atherosclerosis, Alzheimer's disease, rheumatoid arthritis, and chronic heart failure. The most common non-cross linking AGE, carboxymethyl lysine (CML) is formed by the oxidative breakdown of fructosyllysine, which is a product of glucose and lysine. CML is formed in a wide variety of tissues and is an index to assess the extent of glycoxidative damage. Thus we have constructed a mathematical and computational model that predicts the effect of temperature differences in vivo, on the formation of CML, which is now being considered as an important intracellular milieu. This hybrid model that had been tested for its parameter fitting and its sensitivity with available experimental data paves the way for designing novel laboratory experiments that would throw more light on the pathological formation of AGE adducts and in the pathophysiology of diabetic complications.

Keywords: advanced glycation end-products, CML, mathematical model, computational model

Procedia PDF Downloads 126
622 Effect of Feeding Broilers on Diets Enriching With Omega-3 Fatty Acids Sources

Authors: Khalid Mahmoud Gaafar

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In human diets , ω-6 and ω-3 are important essential fatty acids for immunity and health. However, considerable alteration in dietary patterns and contents has resulted in change of the consumption of such fatty acids ,with subsequent increase in the consumption of ω-6 fatty acids and a marked decrease in the consumption of ω-3 fatty acids. This dietary alteration has led to an imbalance in the ratio for ω-6/ω-3, which at 20:1 now differs considerably from the original ratio (1:1). Therefore, dietary supplements such as eggs and meat enriched with omega 3 are necessary to increase the consumption of ω-3 to meet the recommended need for ω-3. Foods that supply ω-6 fatty acids include soybean, palm , sunflower, and rapeseed oils, whereas foods that supply ω-3 fatty acids such as linseed and fish oils. Lin seed oils contain Alpha – linolenic acid (ALA), which can be converted to DHA and EPA in the birds body, with linseed oil containing more than 50% ALA. On the other hand, high doses of omega 6 sources in the diet may have deleterious effects on humans. Maintaining an optimum ratio of ω-3 and ω-6fatty acids not only improves performance but also prevents these health risks. The ratio of n-6:ω-3 fatty acids also plays an important role in the immune response, production performance of broilers and designing meat enriched with ω-3 polyunsaturated fatty acids (PUFAs). Birds of three experimental groups fed on basal starter (0-2nd weeks), grower (3rd -4th weeks) and finisher (5th week) rations. The first is control group fed during the grower-finisher periods on basic diet with two replicate (one fed on basic diet contain vegetable oil and the other don’t) without any additives. The three experimental groups (T1 – T2 –T3) fed during the grower- finisher periods on diets free from vegetable oils and contain of 5% of extruded mixture of soybean and linseed (60%:40%). The second (T2) and third (T3) experimental groups supplemented with vitamin B12 and enzyme mixture. The first experimental groups don’t receive vitamins or enzymes. The obtained results showed a significant increased growth performance, immune response, highest antioxidant activity and serum HDL with lowest serum LDL and triglycerides levels in all experimental groups compared with control group, which was highly significant in group fed on vitamin B6.

Keywords: omega fatty acids, broiler, feeding, human health, growth performance, immunity

Procedia PDF Downloads 101
621 The Relationships between AntimüLlerian Hormone, Androgens and Ovarian Reserve in Non-Obese East Indian Women with and without Polycystic Ovary Syndrome

Authors: Dipanshu Sur, Ratnabali Chakravorty, Rimi Pal, Siddhartha Chatterjee, Joyshree Chaterjee, Amal Mallik

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Background: Polycystic ovary syndrome (PCOS) is a common endocrine disease in reproductive women with a complex hormonal disturbance that affects the menstrual cycle and leads to metabolic consequences in later life. Hyperandrogenaemia is noticeable features of PCOS and influence the process of folliculogenesis in women. The levels of Antimüllerian Hormone (AMH) reflect the number of pre-antral follicles and thus are a marker of oocyte pool – germinal reserve of the ovary for reproduction. Besides its utilization in IVF (In-vitro fertilization), determination of AMH may serve as an additional marker in the diagnostics of PCOS, where increased AMH levels reflect the severity of the disease. The positive correlation of serum AMH with the number of antral follicles was found also in patients with PCOS. Objective: The objective of this study was to investigate the relationship between AMH androgens and whether AMH contributes to altered folliculogenesis in non-obese women with PCOS. Methods: We designed a prospective study which included a total of 65 IVF individuals. It enrolled 26 cases of PCOS based on 2003 Rotterdam criteria and 39 ovulatory normal- non PCOS, healthy, age-matched controls. AMH levels and ovarian morphology were assessed. The relationships between AMH and androgenaemia in patients with and without PCOS were studied. Results: Mean age of PCOS patients were slightly higher than controls (32±4 and 28±3 years, respectively). AMH generally increased with antral follicle count (AFC) [P=0.001], testosterone, and luteinising hormone, and decreased with age, and serum sex hormone binding globulin (SHBG). No significant relationships were found between circulating AMH levels and BMI between PCOS and non-PCOS patients. The calculation of AMH production per antral follicle (AMH/AF) showed that there was a significant difference in median AMH/AF between PCOS and non-PCOS (P =0.001). Both PCOS and non-PCOS groups showed a very similar increase in AMH with increases in AFC, but the PCOS patients had consistently higher AMH across all AFC levels. Conclusions: These observations indicate that there is a connection between AMH and androgens levels between PCOS and non-PCOS East Indian women. Excessive granulosa cell activity may be implicated in the abnormal follicular dynamic of the syndrome. They are higher in women with PCOS and, on the other hand, very low in women with an ovarian failure.

Keywords: anti-Mullerian hormone, polycystic ovary syndrome, antral follicle count, androgens

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620 Oil Extraction from Sunflower Seed Using Green Solvent 2-Methyltetrahydrofuran and Isoamyl Alcohol

Authors: Sergio S. De Jesus, Aline Santana, Rubens Maciel Filho

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The objective of this study was to choose and determine a green solvent system with similar extraction efficiencies as the traditional Bligh and Dyer method. Sunflower seed oil was extracted using Bligh and Dyer method with 2-methyltetrahydrofuran and isoamyl using alcohol ratios of 1:1; 2:1; 3:1; 1:2; 3:1. At the same time comparative experiments was performed with chloroform and methanol ratios of 1:1; 2:1; 3:1; 1:2; 3:1. Comparison study was done using 5 replicates (n=5). Statistical analysis was performed using Microsoft Office Excel (Microsoft, USA) to determine means and Tukey’s Honestly Significant Difference test for comparison between treatments (α = 0.05). The results showed that using classic method with methanol and chloroform presented the extraction oil yield with the values of 31-44% (w/w) and values of 36-45% (w/w) using green solvents for extractions. Among the two extraction methods, 2 methyltetrahydrofuran and isoamyl alcohol ratio 2:1 provided the best results (45% w/w), while the classic method using chloroform and methanol with ratio of 3:1 presented a extraction oil yield of 44% (w/w). It was concluded that the proposed extraction method using 2-methyltetrahydrofuran and isoamyl alcohol in this work allowed the same efficiency level as chloroform and methanol.

Keywords: extraction, green solvent, lipids, sugarcane

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619 Development and Evaluation of a Calcium Rich Plant-Based Supplement on Bone Turnover of Peri and Post Menopausal Women

Authors: Gayathri.G, Hemamalini.A.J, Chandrasekaran.A

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Problem statement: Nutritional deficiency, especially calcium, may lead to poor bone formation and mineralization. Although there are plenty of synthetic supplements available, it is essential to make a calcium rich food supplement accessible to combat calcium deficiency that could be readily prepared at the household level. Thus the current study aimed to formulate and standardize an indigenous low-cost calcium-rich food supplement and to study the impact of supplementation on the bone resorption and formation markers. Methods: A Randomized controlled trial was conducted with 60 subjects distributed equally in control and experimental groups, including perimenopausal and postmenopausal women. A plant-based calcium-rich product was developed and supplemented in form of balls as a midmorning and evening snack by addition of optimized proportions of leaves of Sesbania Grandiflora, seeds of Sesamum indicum, Eleusine coracana, Glycine max, Vigna mungo for a period of 6 months. Postmenopausal and perimenopausal women received 1200mg and 800mg of calcium per day from the supplemented, respectively. Outcome measures like serum calcium; betacrosslaps (bone resorption marker) and total P1NP (bone absorption marker) were assessed after 3 months and after 6 months. Results: There were no significant changes seen in the serum calcium and total P1NP levels (bone formation marker) among the subjects during the supplementation period. The bone resorption marker (betacrosslaps) reduced in all the groups and the reduction (0.32 ± 0.130 ng/ml to 0.25 ± 0.130 ng/ml) was found to be statistically highly significant (p < 0.01) in experimental group of perimenopausal subjects and significant (p < 0.05) in experimental group of postmenopausal subjects (1.11 ± 0.290 ng/ml to 0.42 ± 0.263 ng/ml). Conclusion: With the current severe calcium deficiency in the Indian population, integrating low-cost, calcium-rich native foods that could be readily prepared at household level would be useful in raising the nutritional consumption of calcium, which would, in turn, decrease bone turnover.

Keywords: calcium, sesbania grandiflora, sesamum indicum, eleusine coracana, glycine max, vigna mungo, postmenopause, perimenopause, bone resorption, bone absorption, betacrosslaps, total P1NP

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618 Fine Characterization of Glucose Modified Human Serum Albumin by Different Biophysical and Biochemical Techniques at a Range

Authors: Neelofar, Khursheed Alam, Jamal Ahmad

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Protein modification in diabetes mellitus may lead to early glycation products (EGPs) or amadori product as well as advanced glycation end products (AGEs). Early glycation involves the reaction of glucose with N-terminal and lysyl side chain amino groups to form Schiff’s base which undergoes rearrangements to form more stable early glycation product known as Amadori product. After Amadori, the reactions become more complicated leading to the formation of advanced glycation end products (AGEs) that interact with various AGE receptors, thereby playing an important role in the long-term complications of diabetes. Millard reaction or nonenzymatic glycation reaction accelerate in diabetes due to hyperglycation and alter serum protein’s structure, their normal functions that lead micro and macro vascular complications in diabetic patients. In this study, Human Serum Albumin (HSA) with a constant concentration was incubated with different concentrations of glucose at 370C for a week. At 4th day, Amadori product was formed that was confirmed by colorimetric method NBT assay and TBA assay which both are authenticate early glycation product. Conformational changes in native as well as all samples of Amadori albumin with different concentrations of glucose were investigated by various biophysical and biochemical techniques. Main biophysical techniques hyperchromacity, quenching of fluorescence intensity, FTIR, CD and SDS-PAGE were used. Further conformational changes were observed by biochemical assays mainly HMF formation, fructoseamine, reduction of fructoseamine with NaBH4, carbonyl content estimation, lysine and arginine residues estimation, ANS binding property and thiol group estimation. This study find structural and biochemical changes in Amadori modified HSA with normal to hyperchronic range of glucose with respect to native HSA. When glucose concentration was increased from normal to chronic range biochemical and structural changes also increased. Highest alteration in secondary and tertiary structure and conformation in glycated HSA was observed at the hyperchronic concentration (75mM) of glucose. Although it has been found that Amadori modified proteins is also involved in secondary complications of diabetes as AGEs but very few studies have been done to analyze the conformational changes in Amadori modified proteins due to early glycation. Most of the studies were found on the structural changes in Amadori protein at a particular glucose concentration but no study was found to compare the biophysical and biochemical changes in HSA due to early glycation with a range of glucose concentration at a constant incubation time. So this study provide the information about the biochemical and biophysical changes occur in Amadori modified albumin at a range of glucose normal to chronic in diabetes. Although many implicates currently in use i.e. glycaemic control, insulin treatment and other chemical therapies that can control many aspects of diabetes. However, even with intensive use of current antidiabetic agents more than 50 % of diabetic patient’s type 2 suffers poor glycaemic control and 18 % develop serious complications within six years of diagnosis. Experimental evidence related to diabetes suggests that preventing the nonenzymatic glycation of relevant proteins or blocking their biological effects might beneficially influence the evolution of vascular complications in diabetic patients or quantization of amadori adduct of HSA by authentic antibodies against HSA-EGPs can be used as marker for early detection of the initiation/progression of secondary complications of diabetes. So this research work may be helpful for the same.

Keywords: diabetes mellitus, glycation, albumin, amadori, biophysical and biochemical techniques

Procedia PDF Downloads 264
617 Gold Nanoparticle Conjugated with Andrographolide Ameliorates Viper Venom-Induced Inflammatory Response and Organ Toxicity in Animal Model

Authors: Sourav Ghosh, Antony Gomes

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Since 1894 anti-snake venom serum (ASVS) is the only available treatment against snake envenomation, although there are many side effects and limitations. The need for a supportive treatment was felt for a long time to overcome the side effects and limitations of ASVS. Andrographolide conjugated with gold nanoparticle (A-GNP) has been found to antagonize viper venom-induced local damages. The present study was aimed to study the protective efficacy of A-GNP against Viper venom-induced inflammatory response and organ toxicity in animal model. Ethical clearance was obtained from animal experiments. Physico-chemical characterization of A-GNP was done by DLS (diameter and zeta potential), FE-SEM and XRD. Swiss albino male mice were divided into 4 groups: Gr.1-Sham control, Gr.2- Russell’s Viper venom (RVV) control, Gr.3- andrographolide treated and Gr.4- A-GNP treated. The 1/5th minimum lethal dose of RVV (500µg/kg, s.c.) was induced in animals of group 2, 3 & 4 animals, followed by treatment with andrographolide (100mg/kg, i.p.) and A-GNP (100mg/kg, i.v.) in group 3 & 4 animals, respectively. Blood was collected after 18 h, serum was prepared, and inflammatory markers (IL 1β, 6, 17a, 10, TNF α) and biochemical markers (AST, ACP, LDH, urea, creatinine) were assessed. Values were expressed as mean±SEM (n=4), one way ANOVA was done, P<0.05 was considered as statistically significant. DLS size showed the hydrodynamic diameter of A-GNP to be 230-260nm with polydispersity index of 0.103 and zeta potential was -18.32mV. XRD data confirmed the presence of crystalline gold in A-GNP, and FESEM indicated the presence of nearly spherical particle with size18-24nm.Treatment with A-GNP significantly decreased viper venom-induced proinflammatory markers (IL 1β, 6, 17, TNF α) increased anti-inflammatory markers (IL 10) and decreased organ toxicity markers (AST, ACP, LDH, urea, creatinine) in animal model. Venom neutralization efficacy of A-GNP was > andrographolide, which confirmed the increased efficacy of andrographolide after gold nanoparticle conjugation. Venom neutralization by A-GNP was due to anti-oxidant/anti-inflammatory activity of andrographolide, which showed increased efficacy after gold nanoparticle tagging. Thus, A-GNP may serve as a supportive therapy in snake-bite (against inflammatory response and organ toxicity) subject to further detail studies.

Keywords: andrographolide, gold nanoparticle, inflammatory response, organ toxicity, snake venom, snake venom neutralization, viper venom

Procedia PDF Downloads 363
616 A Radiographic Survey of Eggshell Powder Effect on Tibial Bone Defect Repair Tested in Dog

Authors: M. Yadegari, M. Nourbakhsh, N. Arbabzadeh

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The skeletal system injuries are of major importance. In addition, it is recommended to use materials for hard tissue repair in open or closed fractures. It is important to use complex minerals with a beneficial effect on hard tissue repair, stimulating cell growth in the bone. Materials that could help avoid bone fracture inflammatory reaction and speed up bone fracture repair are of utmost importance in the treatment of bone fractures. Similar to minerals, the inner eggshell membrane consists of carbohydrates, lipids, proteins with the high pH, high calcium absorptive capacity and with faster bone fracture repair ability. In the present radiographic survey, eggshell-derived bone graft substitutes were used for bone defect repair in 8 dog tibia, measuring bone density on the day of implant placement and 30 and 60 days after placement. In fact, the result of this study shows the difference in bone growth and misshapen bones between treatment and control sites. Cell growth was adequate in treatment sites and misshapen bones were less frequent here than in control sites.

Keywords: bone repair, eggshell powder, implant, radiography

Procedia PDF Downloads 313
615 Biodiesel Production from Fruit Pulp of Cassia fistula L. Using Green Microalga Chlorella minutissima

Authors: Rajesh Chandra, Uttam K. Ghosh

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This study demonstrates microalgal bio-diesel generation from a cheap, abundant, non-edible fruit pulp of Cassia fistula L. The Cassia fistula L. fruit pulp aqueous extract (CFAE) was utilized as a growth medium for cultivation of microalga Chlorella minutissima (C. minutissima). This microalga accumulated a high amount of lipids when cultivated with CFAE as a source of nutrition in comparison to BG-11 medium. Different concentrations (10, 20, 30, 40 and 50%) of CFAE diluted with distilled water were used to cultivate microalga. Effects of light intensity and photoperiod were also observed on biomass and lipid yield of microalga. Light intensity of 8000 lux with a photoperiod of 18 h resulted in maximum biomass and lipid yield of 1.28 ± 0.03 and 0.3968 ± 0.05 g/L, respectively when cultivated with 40% CFAE. Fatty acid methyl ester (FAME) profile of bio-diesel obtained shown the presence of myristic acid (C14:0), palmitic acid (C16:0), palmitoleic acid (C16:1), stearic acid (C18:0), linoleic acid (C18:2), linolenic acid (C18:3), arachidic acid (C20:0), and gondoic acid (C20:1), as major fatty acids. These facts reflect that the fruit pulp of Cassia fistula L. can be used for cultivation of C. minutissima.

Keywords: biomass, bio-diesel, Cassia fistula L., C. minutissima, GC-MS, lipid

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614 Prednisone and Its Active Metabolite Prednisolone Attenuate Lipid Accumulation in Macrophages

Authors: H. Jeries, N. Volkova, C. G. Iglesias, M. Najjar, M. Rosenblat, M. Aviram, T. Hayek

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Background: Synthetic forms of glucocorticoids (e.g., prednisone, prednisolone) are anti-inflammatory drugs which are widely used in clinical practice. The role of glucocorticoids (GCs) in cardiovascular diseases including atherosclerosis is highly controversial, and their impact on macrophage foam cell formation is still unknown. Our aim was to investigate the effects of prednisone or its active metabolite, prednisolone, on macrophage oxidative stress and lipid metabolism using in-vivo, ex-vivo and in-vitro systems. Methods: The in-vivo study included C57BL/6 mice which were intraperitoneally injected with prednisone or prednisolone (5mg/kg) for 4 weeks, followed by lipid metabolism analyses in the mice aorta, and in peritoneal macrophages (MPM). In the ex-vivo study, we analyzed the effect of serum samples obtained from 9 healthy volunteers before or after treatment with oral prednisone (20mg for 5 days), on J774A.1 macrophage atherogenicity. In-vitro studies were conducted using J774A.1 macrophages, human monocyte derived macrophages (HMDM) and fibroblasts. Cells were incubated with increasing concentrations (0-200 ng/ml) of prednisone or prednisolone, followed by determination of cellular oxidative status, triglyceride and cholesterol metabolism. Results: Prednisone or prednisolone treatment resulted in a significant reduction in triglycerides and mainly in cholesterol cellular accumulation in MPM or in J774A.1 macrophages incubated with human serum. Similar resulted were noted in HMDM or in J774A.1 macrophages which were directly incubated with the GCs. These effects were associated with GCs inhibitory effect on triglycerides and cholesterol biosynthesis rates, throughout downregulation of diacylglycerol acyltransferase1 (DGAT1) expression, and of the sterol regulatory element binding protein (SREBP2) and HMGCR expression, respectively. In parallel to prednisone or prednisolone induced reduction in macrophage triglyceride content, paraoxonase 2 (PON2) expression was significantly upregulated. GCs-induced reduction of cellular triglyceride and cholesterol mass was mediated by the GCs receptors on macrophages since the GCs receptor antagonist (RU 486) abolished these effects. In fibroblasts, unlike macrophages, prednisone or prednisolone showed no anti-atherogenic effects. Conclusions: Prednisone or prednisolone are anti-atherogenic since they protected macrophages from lipid accumulation and foam cell formation.

Keywords: atherosclerosis, cholesterol, foam cell, macrophage, prednisone, prednisolone, triglycerides

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613 Assessment of Cassava Varieties in Ecuador for the Production of Lactic Acid From Starch by-Products

Authors: Pedro Maldonado-Alvarado

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An important cassava quality production was detected in Ecuador. However, in this country, few products with low adding-value are produced from the tuber and none from cassava by-products. To our best knowledge, lactic acid was produced from Ecuadorian cassava bagasse starch in a biotechnological way. The objective of this contribution was to study the influence of the fermentation variables (pH and agitation) on the lactic acid production of Ecuadorian cassava varieties from bagasse starch. Enzymatic hydrolysis of cassava bagasse starch for INIAP 650 and INIAP 651 varieties spread in Ecuador was performed using α-amylase and amyloglucosidase. Then, glucose was fermented by Lactobacillus leichmannii strains in different conditions of agitation (0 and 150 rpm) and pH (4.5, 5.0, and 5.5). Significant differences in ash, fibre, protein, lipids, and amylose were found in cassava bagasse starch of INIAP 650 and INIAP 651 with 1.4 and 1.3%, 4.3 and 6%, 1.2 and 2.1%, 1.9 and 1.5%, and 24.3 and 26.5%, respectively. The determination of lactic acid was performed by potentiometric and FTIR analysis. Conversions of cassava bagasse to reduced sugars were 71.7 and 85.1% for INIAP 650 and INIAP 651, respectively. The best lactic acid concentrations were 27.6 and 33.5 g/L, obtained at agitation 150 rpm and pH 5.5 for INIAP 650 and INIAP 651. Qualitative analysis conducted by FTIR spectrophotometry confirmed the presence of lactic acid in the reacted products. This investigation could contribute to the valorisation of residues from promising cassava varieties in Ecuador and hence to increase the development of this country.

Keywords: bagasse starch, cassava, Ecuador, fermentation, lactic acid

Procedia PDF Downloads 188
612 Study of Oxidative Processes in Blood Serum in Patients with Arterial Hypertension

Authors: Laura M. Hovsepyan, Gayane S. Ghazaryan, Hasmik V. Zanginyan

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Hypertension (HD) is the most common cardiovascular pathology that causes disability and mortality in the working population. Most often, heart failure (HF), which is based on myocardial remodeling, leads to death in hypertension. Recently, endothelial dysfunction (EDF) or a violation of the functional state of the vascular endothelium has been assigned a significant role in the structural changes in the myocardium and the occurrence of heart failure in patients with hypertension. It has now been established that tissues affected by inflammation form increased amounts of superoxide radical and NO, which play a significant role in the development and pathogenesis of various pathologies. They mediate inflammation, modify proteins and damage nucleic acids. The aim of this work was to study the processes of oxidative modification of proteins (OMP) and the production of nitric oxide in hypertension. In the experimental work, the blood of 30 donors and 33 patients with hypertension was used. For the quantitative determination of OMP products, the based on the reaction of the interaction of oxidized amino acid residues of proteins and 2,4-dinitrophenylhydrazine (DNPH) with the formation of 2,4-dinitrophenylhydrazones, the amount of which was determined spectrophotometrically. The optical density of the formed carbonyl derivatives of dinitrophenylhydrazones was recorded at different wavelengths: 356 nm - aliphatic ketone dinitrophenylhydrazones (KDNPH) of neutral character; 370 nm - aliphatic aldehyde dinirophenylhydrazones (ADNPH) of neutral character; 430 nm - aliphatic KDNFG of the main character; 530 nm - basic aliphatic ADNPH. Nitric oxide was determined by photometry using Grace's solution. Adsorption was measured on a Thermo Scientific Evolution 201 SF at a wavelength of 546 nm. Thus, the results of the studies showed that in patients with arterial hypertension, an increased level of nitric oxide in the blood serum is observed, and there is also a tendency to an increase in the intensity of oxidative modification of proteins at a wavelength of 270 nm and 363 nm, which indicates a statistically significant increase in aliphatic aldehyde and ketone dinitrophenylhydrazones. The increase in the intensity of oxidative modification of blood plasma proteins in the studied patients, revealed by us, actually reflects the general direction of free radical processes and, in particular, the oxidation of proteins throughout the body. A decrease in the activity of the antioxidant system also leads to a violation of protein metabolism. The most important consequence of the oxidative modification of proteins is the inactivation of enzymes.

Keywords: hypertension (HD), oxidative modification of proteins (OMP), nitric oxide (NO), oxidative stress

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