Search results for: serum enzymes

Authors: F. Alaa Ali, S. A. Sherein Abdelgayed, S. Osama. EL-Tawil, M. Adel Bakeer

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Tartrazine is an organic azo dyes food additive widely used in foods, drugs, and cosmetics. The present study aimed to investigate the toxic effects of tartrazine on kidneys and liver biomarkers in addition to the investigation of oxidative stress and change of histopathological structure of liver and kidneys in 30 male rats. Tartrazine was orally administrated daily at dose 200 mg/ kg bw (1/ 10 LD₅₀) for sixty days. Serum and tissue samples were collected at the end of the experiment to investigate the underlying mechanism of tartrazine through assessment oxidative stress (Glutathione (GSH), Superoxide dismutase (SOD) and malondialdehyde (MDA) and biochemical markers (alanine aminotransferase (ALT), aspartate aminotransferase (AST), Total protein and Urea). Liver and kidneys tissue were collected and preserved in 10% formalin for histopathological examination. The obtained values were statistically analyzed by one way analysis of variance (ANOVA) followed by multiple comparison test. Biochemical analysis revealed that tartrazine induced significant increase in serum ALT, AST, total protein, urea level compared to control group. Tartrazine showed significant decrease in liver GSH and SOD where their values when compared to control group. Tartrazine induced increase in liver MDA compared to control group. Histopathology of the liver showed diffuse vacuolar degeneration in hepatic parenchyma, the portal area showed sever changes sever in hepatoportal blood vessels and in the bile ducts. The kidneys showed degenerated tubules at the cortex together with mononuclear leucocytes inflammatory cells infiltration. There is perivascular edema with inflammatory cell infiltration surrounding the congested and hyalinized vascular wall of blood vessel. The present study indicates that the subchronic effects of tartrazine have a toxic effect on the liver and kidneys together with induction of oxidative stress by formation of free radicals. Therefore, people should avoid the hazards of consuming tartrazine.

Keywords: albino rats, tartrazine, toxicity, pathology

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Authors: Frans J. Smit, Wisdom A. Munzeiwa, Hermanus C. M. Vosloo, Lyn-Marie Birkholtz, Richard K. Haynes

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Malaria and antimicrobial infections remain significant global health concerns, necessitating the continuous search for novel therapeutic approaches. This abstract presents an overview of the potential use of triazenes as effective agents against malaria and various antimicrobial pathogens. Triazenes are a class of compounds characterized by a linear arrangement of three nitrogen atoms, rendering them structurally distinct from their cyclic counterparts. This study investigates the efficacy of triazenes against malaria and explores their antimicrobial activity. Preliminary results revealed significant antimalarial activity of the triazenes, as evidenced by in vitro screening against P. falciparum, the causative agent of malaria. Furthermore, the compounds exhibited broad-spectrum antimicrobial activity, indicating their potential as effective antimicrobial agents. These compounds have shown inhibitory effects on various essential enzymes and processes involved in parasite survival, replication, and transmission. The mechanism of action of triazenes against malaria involves interactions with critical molecular targets, such as enzymes involved in the parasite's metabolic pathways and proteins responsible for host cell invasion. The antimicrobial activity of the triazenes against bacteria and fungi was investigated through disc diffusion screening. The antimicrobial efficacy of triazenes has been observed against both Gram-positive and Gram-negative bacteria, as well as multidrug-resistant strains, making them potential candidates for combating drug-resistant infections. Furthermore, triazenes possess favourable physicochemical properties, such as good stability, solubility, and low toxicity, which are essential for drug development. The structural versatility of triazenes allows for the modification of their chemical composition to enhance their potency, selectivity, and pharmacokinetic properties. These modifications can be tailored to target specific pathogens, increasing the potential for personalized treatment strategies. In conclusion, this study highlights the potential of triazenes as promising candidates for the development of novel antimalarial and antimicrobial therapeutics. Further investigations are necessary to determine the structure-activity relationships and optimize the pharmacological properties of these compounds. The results warrant additional research, including MIC studies, to further explore the antimicrobial activity of the triazenes. Ultimately, these findings contribute to the development of more effective strategies for combating malaria and microbial infections.

Keywords: malaria, anti-microbials, triazene, resistance

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Authors: Elnaz Saeedi, Jamileh Abolaghasemi, Mohsen Nasiri Tousi, Saeedeh Khosravi

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Goals and Objectives: A typical analysis of survival data involves the modeling of time-to-event data, such as the time till death. A frailty model is a random effect model for time-to-event data, where the random effect has a multiplicative influence on the baseline hazard function. This article aims to investigate the use of gamma frailty model with concomitant variable in order to individualize the prognostic factors that influence the liver cirrhosis patients’ survival times. Methods: During the one-year study period (May 2008-May 2009), data have been used from the recorded information of patients with liver cirrhosis who were scheduled for liver transplantation and were followed up for at least seven years in Imam Khomeini Hospital in Iran. In order to determine the effective factors for cirrhotic patients’ survival in the presence of latent variables, the gamma frailty distribution has been applied. In this article, it was considering the parametric model, such as Exponential and Weibull distributions for survival time. Data analysis is performed using R software, and the error level of 0.05 was considered for all tests. Results: 305 patients with liver cirrhosis including 180 (59%) men and 125 (41%) women were studied. The age average of patients was 39.8 years. At the end of the study, 82 (26%) patients died, among them 48 (58%) were men and 34 (42%) women. The main cause of liver cirrhosis was found hepatitis 'B' with 23%, followed by cryptogenic with 22.6% were identified as the second factor. Generally, 7-year’s survival was 28.44 months, for dead patients and for censoring was 19.33 and 31.79 months, respectively. Using multi-parametric survival models of progressive and regressive, Exponential and Weibull models with regard to the gamma frailty distribution were fitted to the cirrhosis data. In both models, factors including, age, bilirubin serum, albumin serum, and encephalopathy had a significant effect on survival time of cirrhotic patients. Conclusion: To investigate the effective factors for the time of patients’ death with liver cirrhosis in the presence of latent variables, gamma frailty model with parametric distributions seems desirable.

Keywords: frailty model, latent variables, liver cirrhosis, parametric distribution

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Authors: Rayhana Begum, Manju Sharma

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Careya arborea Roxb. belongs to the Lecythidaceae family, is traditionally used in tumors, anthelmintic, bronchitis, epileptic fits, astringents, inflammation, an antidote to snake-venom, skin disease, diarrhea, dysentery with bloody stools, dyspepsia, ulcer, toothache, and ear pain. The present study was focused on investigating the anti-arthritic effect of coumaroyl lupendioic acid, a new lupane-type triterpene from Careya arborea stem bark in the chronic inflammatory model and further assessing its possible mechanism on the modulation of inflammatory biomarkers. Arthritis was induced by injecting 0.1 ml of Complete Freund’s Adjuvant (5 mg/ml of heat killed Mycobacterium tuberculosis) into the subplantar region of the left hind paw. Treatment with coumaroyl lupendioic acid (10 and 20 mg/kg, p.o.) and reference drugs (indomethacin and dexamethasone at the dose of 5 mg/kg, p.o.) were started on the day of induction and continued up to 28 days. The progression of arthritis was evaluated by measuring paw volume, tibio tarsal joint diameters, and arthritic index. The effect of coumaroyl lupendioic acid (CLA) on the production PGE₂, NO, MPO, NF-κB, TNF-α, IL-1β, and IL-6 on serum level as well as inflamed paw tissue were also assessed. In addition, ankle joints and spleen were collected and prepared for histological examination. CLA in inflamed rats resulted in significant amelioration of paw edema, tibio-tarsal joint swelling and arthritic score as compared to CFA control group. The results indicated that CLA treated groups markedly decreased the levels of inflammatory mediators (PGE₂, NO, MPO and NF-κB levels) and down-regulated the production of pro-inflammatory cytokines (TNF-α, IL-1β, and IL-6) in paw tissue homogenates as well as in serum. However, the more pronounced effect was observed in the inflamed paw tissue homogenates. CLA also revealed a protective effect to the tibio-tarsal joint cartilage and spleen. These results suggest that coumaroyl lupendioic acid inhibits inflammation may be through the suppression of the cascade of proinflammatory mediators via the down-regulation of NF-ҡB.

Keywords: complete Freund’s adjuvant , Coumaroyl lupendioic acid, pro-inflammatory cytokines, prostaglandin E2

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Authors: Muhammad Ajmal

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- Thermal processing and subsequent storage of ultra-heat treated (UHT) milk leads to alteration in protein profile, Maillard reaction and lipid oxidation. Concentration of carbohydrates in normal and flavored version of UHT milk is considerably different. Transition in protein profile, Maillard reaction and lipid oxidation in UHT flavored milk was determined for 90 days at ambient conditions and analyzed at 0, 45 and 90 days of storage. Protein profile, hydroxymethyl furfural, furosine, Nε-carboxymethyl-l-lysine, fatty acid profile, free fatty acids, peroxide value and sensory characteristics were determined. After 90 days of storage, fat, protein, total solids contents and pH were significantly less than the initial values determined at 0 day. As compared to protein profile normal UHT milk, more pronounced changes were recorded in different fractions of protein in UHT milk at 45 and 90 days of storage. Tyrosine content of flavored UHT milk at 0, 45 and 90 days of storage were 3.5, 6.9 and 15.2 µg tyrosine/ml. After 45 days of storage, the decline in αs1-casein, αs2-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, immunoglobulin and bovine serum albumin were 3.35%, 10.5%, 7.89%, 18.8%, 53.6%, 20.1%, 26.9 and 37.5%. After 90 days of storage, the decline in αs1-casein, αs2-casein, β-casein, κ-casein, β-lactoglobulin, α-lactalbumin, immunoglobulin and bovine serum albumin were 11.2%, 34.8%, 14.3%, 33.9%, 56.9%, 24.8%, 36.5% and 43.1%. Hydroxy methyl furfural content of UHT milk at 0, 45 and 90 days of storage were 1.56, 4.18 and 7.61 (µmol/L). Furosine content of flavored UHT milk at 0, 45 and 90 days of storage intervals were 278, 392 and 561 mg/100g protein. Nε-carboxymethyl-l-lysine content of UHT flavored milk at 0, 45 and 90 days of storage were 67, 135 and 343mg/kg protein. After 90 days of storage of flavored UHT milk, the loss of unsaturated fatty acids 45.7% from the initial values. At 0, 45 and 90 days of storage, free fatty acids of flavored UHT milk were 0.08%, 0.11% and 0.16% (p<0.05). Peroxide value of flavored UHT milk at 0, 45 and 90 days of storage was 0.22, 0.65 and 2.88 (MeqO²/kg). Sensory analysis of flavored UHT milk after 90 days indicated that appearance, flavor and mouth feel score significantly decreased from the initial values recorded at 0 day. Findings of this investigation evidenced that in flavored UHT milk more pronounced changes take place in protein profile, Maillard reaction products and lipid oxidation as compared to normal UHT milk.

Keywords: UHT flavored milk , hydroxymethyl furfural, lipid oxidation, sensory properties

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Authors: Alexander A. Osmolovskiy, Anastasia V. Orekhova, Daria M. Bednenko, Yelyzaveta Boiko

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Micromycetes from Aspergillus genus can produce proteases capable of promoting proteolysis of hemostasis proteins or, along with hydrolytic activity, to show the ability to convert proenzymes of this system activating them into an active form. At the same time, practical medicine needs specific activators for quantitation of the level of some plasma enzymes, especially protein C and factor X, the lack of which leads to the development of thromboembolic diseases. Thus, some micromycetes of the genus Aspergillus were screened for the ability to synthesize extracellular proteases with promising activity for designing anti-thrombotic and diagnostic preparations. Such standard methods like salting out, electrophoresis, isoelectrofocusing were used for isolation, purification and study of physicochemical properties of proteases. Enzyme activity was measured spectrophotometrically fibrin as a substrate of the reaction and chromogenic peptide substrates of different proteases of the human hemostasis system. As a result of the screening, four active producers were selected: Aspergillus janus 301, A. flavus 1, A. terreus 2, and A. ochraceus L-1. The enzyme of A. janus 301 showed the greatest fibrinolytic activity (around 329.2 μmol Tyr/(ml × min)). The protease produced by A. terreus 2 had the highest plasmin-like activity (54.1 nmol pNA/(ml × min)), but fibrinolytic activity was lower than A. janus 301 demonstrated (25.2 μmol Tyr/(ml × min)). For extracellular protease of micromycete A. flavus a high plasmin-like activity was also shown (39.8 nmol pNA / (ml × min)). Moreover, according to our results proteases one of the fungi - A. terreus 2 were able to activate protein C of human plasma - the key factor of the human anticoagulant hemostasis system. This type of activity was 39.8 nmol pNA/(ml × min)). It was also shown that A. ochraceus L-1 could produce extracellular proteases with protein C and factor X activator activities (65.9 nmol pNA/(ml × min) and 34.6 nmol pNA/(ml × min) respectively). The maximum accumulation of the proteases falls on the 4th day of cultivation. Using isoelectrofocusing was demonstrated that the activation of both proenzymes might proceed via limited proteolysis induced by proteases of A. ochraceus L-1. The activatory activity of A. ochraceus L-1 proteases toward essential hemostatic proenzymes, protein C and X factor may be useful for practical needs. It is well known that similar enzymes, activators of protein C and X factor isolated from snake venom, South American copperhead Agkistrodon contortrix contortrix and Russell’s viper Daboia russelli russeli, respectively, are used for the in vitro diagnostics of the functional state of these proteins in blood plasma. Thus, the proteases of Aspergillus genus can be used as cheap components for enzyme thrombolytic preparations.

Keywords: anti-trombotic drugs, fibrinolysis, diagnostics, proteases, micromycetes

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Authors: Mardiana Madjid, Nurpudji Astuti Taslim, Suryani As'ad, Haerani Rasyid, Agussalim Bukhari

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The high level of Urine Urea Nitrogen (UUN) indicates hypercatabolism occurs in hospitalized patients. High levels of Total Lymphocyte Count (TLC) indicates the immune system condition, adequate wound healing, and limit complication. Adequate dietary intake affects to decrease of hypercatabolism status in treated patient’s hospitals. Nitrogen Balance (NB) is simply the difference between nitrogen (N₂) intake and output. If more N₂ intake than output, then positive NB or anabolic will occur. This study aims to evaluate the effect of dietary intake in influencing balance nitrogen and total lymphocyte count. Method: A total of 43 patients admitted to a Wahidin Sudirohusodo Hospital between 2018 and 2019 for 10 days' treats are included. The inclusion criteria were patients who were treated for 10 days and receives food from the hospital orally. Patients did not experience gastrointestinal disorders such as vomiting and diarrhea and experience impair kidney function and liver function and expressed approval to participate in this study. During hospitalization, food intake, UUN, albumin serum, balance nitrogen, and TLC was assessed twice on day 1 and day 10. There is no Physician Clinical Nutritional intervention to correct food intake. UUN is 24 hours of urine collected on the second day after admission and the tenth day. Statistical analysis uses SPSS 24 with observational cohort methods. Result: The Forty-three participants completed the follow-up (27 men and 18 women). The age of fewer than 4 years is 22 people, 45 to 60 years is 16 people, and over 60 years is 4 people. The result of the study on day 1 obtained SGA score A, SGA score B, SGA score C are 8, 32, 3 until day 10 are 8, 31, 4, respectively. According to 24h dietary recalls, the energy intake during observation was from 522.5 ± 400.4 to 1011.9 ± 545.1 kcal/day P < 0.05, protein intake from 20.07 ± 17.2 to 40.3 ± 27.3 g/day P < 0.05, carbohydrates from 92.5 ± 71.6 to 184.8 ± 87.4 g/day, and fat from 5.5 ± 3.86 to 13.9 ± 13.9 g/day. The UUN during the observation was from 6.6 ± 7.3 to 5.5 ± 3.9 g/day, TLC decreased from 1622.9 ± 897.2 to 1319.9 ± 636.3/mm³ value target 1800/mm³, albumin serum from 3.07 ± 0.76 to 2.9 ± 0.57 g/day, and BN from -7.5 ± 7.2 to -3.1 ± 4.86. Conclusion: The high level of UUN needs to correct adequate dietary intake to improve NB and TLC status on hospitalized patients.

Keywords: adequate dietary intake, balance nitrogen, total lymphocyte count, urine urea nitrogen

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Authors: Abdul-Dahiru El-Yuguda, Saka Saheed Baba, Tawa Monilade Adisa, Mustapha Bala Abubakar

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Background: Chikungunya (CHIK) virus, a previously anecdotally described arbovirus, is now assuming a worldwide public health burden. The CHIK virus infection is characterized by potentially life threatening and debilitating arthritis in addition to the high fever, arthralgia, myalgia, headache and rash. Method: Three hundred and seventy (370) serum samples were collected from outpatients with febrile illness attending University of Maiduguri Teaching Hospital, Nigeria, and was used to detect for Chikungunya (CHIK) virus IgG and IgM antibodies using the Enzyme Linked Immunosorbent Assays (ELISAs). Result: Out of the 370 sera tested, 39 (10.5%) were positive for presence of CHIK virus antibodies. A total of 24 (6.5%) tested positive for CHIK virus IgM only while none (0.0%) was positive for presence of CHIK virus IgG only and 15 (4.1%) of the serum samples were positive for both IgG and IgM antibodies. A significant difference (p<0.0001) was observed in the distribution of CHIK virus antibodies in relation to gender. The males had prevalence of 8.5% IgM antibodies as against 4.6% observed in females. On the other hand 4.6% of the females were positive for concurrent CHIK virus IgG and IgM antibodies when compared to a prevalence of 3.4% observed in males. Only the age groups ≤ 60 years and the undisclosed age group were positive for presence of CHIK virus IgG and/or IgM antibodies. No significant difference (p>0.05) was observed in the seasonal prevalence of CHIK virus antibodies among the study subjects Analysis of the prevalence of CHIK virus antibodies in relation to clinical presentation (as observed by Clinicians) of the patients revealed that headache and fever were the most frequently encountered ailments. Conclusion: The CHIK virus IgM and concurrent IgM and IgG antibody prevalence rates of 6.5% and 4.1% observed in this study indicates a current infection and the lack of IgG antibody alone observed shows that the infection is not endemic but sporadic. Recommendation: Further studies should be carried to establish the seasonal prevalence of CHIK virus infection vis-à-vis vector dynamics in the study area. A comprehensive study need to be carried out on the molecular characterization of the CHIK virus circulating in Nigeria with a view to developing CHIK virus vaccine.

Keywords: Chikungunya virus, IgM and IgG antibodies, febrile patients, enzyme linked immunosorbent assay

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Authors: Eisha Vienna M. Fernandez, Cristan Q. Cabanilla, Hiyasmin Lim, John Donnie A. Ramos

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Allergy is a multifactorial disease affecting a significant proportion of the population. It is developed through the interaction of allergens and the presence of certain polymorphisms in various susceptibility genes. In this study, the correlation of the 105A/C single nucleotide polymorphism (SNP) of the IL-18 gene and house dust mite-specific IgE among Filipino allergic and non-allergic population was investigated. Atopic status was defined by serum total IgE concentration of ≥100 IU/mL, while house dust mite allergy was defined by specific IgE value ≥ +1SD of IgE of nonatopic participants. Two hundred twenty match-paired Filipino cases and controls aged 6-60 were the subjects of this investigation. The level of total IgE and Specific IgE were measured using Enzyme-Linked Immunosorbent Assay (ELISA) while Polymerase Chain Reaction – Restriction Fragment Length Polymorphism (PCR-RFLP) analysis was used in the SNP detection. Sensitization profiles of the allergic patients revealed that 97.3% were sensitized to Blomia tropicalis, 40.0% to Dermatophagoides farinae, and 29.1% to Dermatophagoides pteronyssinus. Multiple sensitization to HDMs was also observed among the 47.27% of the atopic participants. Any of the allergy classes of the atopic triad were exhibited by the cases (allergic asthma: 48.18%; allergic rhinitis: 62.73%; atopic dermatitis: 19.09%), and two or all of these atopic states are concurrently occurring in 26.36% of the cases. A greater proportion of the atopic participants with allergic asthma and allergic rhinitis were sensitized to D. farinae, and D. pteronyssinus, while more of those with atopic dermatitis were sensitized to D. pteronyssinus than D. farinae. Results show that there is overrepresentation of the allele “A” of the 105A/C IL-18 gene SNP in both cases and control groups of the population. The genotype that predominate the population is the heterozygous “AC”, followed by the homozygous wild “AA”, and the homozygous variant “CC” being the least. The study confirmed a positive association between serum specific IgE against B. tropicalis and D. pteronyssinus and the allele “C” (Bt P=0.021, Dp P=0.027) and “AC” (Bt P=0.003, Dp P=0.026) genotype. Findings also revealed that the genotypes “AA” (OR:1.217; 95% CI: 0.701-2.113) and “CC” (OR, 3.5; 95% CI: 0.727-16.849) increase the risk of developing allergy. This indicates that the 105A/C IL-18 gene SNP is a candidate genetic marker for HDM allergy among Filipino patients.

Keywords: house dust mite allergy, interleukin-18 (IL-18), single nucleotide polymorphism,

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

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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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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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Authors: Sujata Sharma, Avinash Singh, Lovely Gautam, Pradeep Sharma, Mau Sinha, Asha Bhushan, Punit Kaur, Tej P. Singh

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Peptidyl-tRNA hydrolase (Pth) Pth is an essential bacterial enzyme that catalyzes the release of free tRNA and peptide moeities from peptidyl tRNAs during stalling of protein synthesis. In order to design inhibitors of Pth from Pseudomonas aeruginosa (PaPth), we have determined the structures of PaPth in its native state and in the bound states with two compounds, amino acylate-tRNA analogue (AAtA) and 5-azacytidine (AZAC). The peptidyl-tRNA hydrolase gene from Pseudomonas aeruginosa was amplified by Phusion High-Fidelity DNA Polymerase using forward and reverse primers, respectively. The E. coliBL21 (λDE3) strain was used for expression of the recombinant peptidyl-tRNA hydrolase from Pseudomonas aeruginosa. The protein was purified using a Ni-NTA superflow column. The crystallization experiments were carried out using hanging drop vapour diffusion method. The crystals diffracted to 1.50 Å resolution. The data were processed using HKL-2000. The polypeptide chain of PaPth consists of 194 amino acid residues from Met1 to Ala194. The centrally located β-structure is surrounded by α-helices from all sides except the side that has entrance to the substrate binding site. The structures of the complexes of PaPth with AAtA and AZAC showed the ligands bound to PaPth in the substrate binding cleft and interacted with protein atoms extensively. The residues that formed intermolecular hydrogen bonds with the atoms of AAtA included Asn12, His22, Asn70, Gly113, Asn116, Ser148, and Glu161 of the symmetry related molecule. The amino acids that were involved in hydrogen bonded interactions in case of AZAC included, His22, Gly113, Asn116, and Ser148. As indicated by fittings of two ligands and the number of interactions made by them with protein atoms, AAtA appears to be a more compatible with the structure of the substrate binding cleft. However, there is a further scope to achieve a better stacking than that of O-tyrosyl moiety because it is not still ideally stacked. These observations about the interactions between the protein and ligands have provided the information about the mode of binding of ligands, nature and number of interactions. This information may be useful for the design of tight inhibitors of Pth enzymes.

Keywords: peptidyl tRNA hydrolase, Acinetobacter baumannii, Pth enzymes, O-tyrosyl

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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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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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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,

Procedia PDF Downloads 115

Authors: Emílie Kučerová, Tereza Veverková, Marina Morozovová, Eva Kudová, Jitka Viktorová

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The blood-brain barrier (BBB) is a key element regulating the transport of substances between the blood and the central nervous system (CNS). The BBB protects the CNS from potentially harmful substances and maintains a suitable environment for nervous activity in the CNS, but at the same time, it represents a significant obstacle to the entry of drugs into the CNS. Pharmacoresistant epilepsy is a form of epilepsy that cannot be suppressed using two (or more) appropriately chosen antiepileptic drugs. In many cases, pharmacoresistant epilepsy is characterized by an increased concentration of efflux pumps on the luminal sides of the endothelial cells that form the BBB and an increased number of drug-metabolizing enzymes in the BBB cells, thereby preventing the effective transport of antiepileptic drugs into the CNS. Currently, a number of scientific groups are focusing on the preparation and improvement of BBB models in vitro in order to study cell interactions or transport mechanisms. However, in pathological conditions such as pharmacoresistant epilepsy, there are changes in BBB structure, and current BBB models are insufficient for related research. Our goal is to develop a suitable BBB model for pharmacoresistant epilepsy in vitro and use it to test the transfer of potential antiepileptic drugs. This model is created by co-culturing immortalized human cerebral microvascular endothelial cells, human vascular pericytes and immortalized human astrocytes. The BBB in vitro is cultivated in the form of a 2D transwell model and the integrity of the barrier is verified by measuring transendothelial electrical resistance (TEER). From the current results, a contact cell arrangement with the cultivation of endothelial cells on the upper side of the insert and the co-cultivation of astrocytes and pericytes on the lower side of the insert is selected as the most promising for BBB model cultivation. The pharmacoresistance of the BBB model is achieved by long-term cultivation of endothelial cells in an increasing concentration of selected antiepileptic drugs, which should lead to increased production of efflux pumps and drug-metabolizing enzymes. The pharmacoresistant BBB model in vitro will be further used for the screening of substances that could act both as antiepileptics and at the same time as inhibitors of efflux pumps in endothelial cells. This project was supported by the Technology Agency of the Czech Republic (TACR), Personalized Medicine: Translational research towards biomedical applications, No. TN02000109 and by the Academy of Sciences of the Czech Republic (AS CR) – grant RVO 61388963.

Keywords: antiepileptic drugs, blood-brain barrier, efflux transporters, pharmacoresistance

Procedia PDF Downloads 53

Authors: Ž. Vaštag, Lj. Popović, S. Popović

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After cold pressing of pumpkin oil, the defatted oil cake (PUOC) was utilized as raw material for processing of bio-functional hydrolysates. In this study, the in vitro bioactivity of an alcalase (AH) and a pepsin hydrolysate (PH) prepared from the major pumpkin 12S globulin (cucurbitin) are compared. The hydrolysates were produced at optimum reaction conditions (temperature, pH) for the enzymes, during 60min. The bioactivity testing included antioxidant and angiotensin I converting enzyme inhibitory activity assays. The hydrolysates showed high potential as natural antioxidants and possibly antihypertensive agents in functional food or nutraceuticals. Additionally, preliminary studies have shown that both hydrolysates could exhibit modest α-amylase inhibitory activity, which indicates on their hypoglycemic potential.

Keywords: cucurbitin, alcalase, pepsin, protein hydrolysates, in vitro bioactivity

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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

Procedia PDF Downloads 394

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

Procedia PDF Downloads 131

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

Procedia PDF Downloads 384

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

Procedia PDF Downloads 148

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

Procedia PDF Downloads 68

Authors: Aruma Baduge Kithma Hansanee De Silva

Abstract:

Haskap (Lonicera caerulea L.), also known as blue honeysuckle, is a recently commercialized berry crop in Canada. Haskap berries are rich in polyphenols, including anthocyanins, which are known for potential health-promoting effects. Cyanidin-3-O-glucoside (C3G) is the most prominent anthocyanin of haskap berries. Recent literature reveals the efficacy of C3G in reducing the risk of type 2 diabetes (T2D), which has become an increasingly common health issue around the world. The T2D is characterized as a metabolic disorder of hyperglycemia and insulin resistance. It has been demonstrated that C3G has anti-diabetic effects in various ways, including improvement in insulin sensitivity, and inhibition of activities of carbohydrate-hydrolyzing enzymes, including alpha-amylase and alpha-glucosidase. The goal of this study was to investigate the influence of variety and harvesting date on haskap composition, biological properties, and antidiabetic properties. The polyphenolic compounds present in four commercially grown haskap cultivars, Aurora, Rebecca, Larissa and Evie among five harvesting stages (H1-H5), were extracted separately in 80% ethanol and analyzed to characterize their phenolic profiles. The haskap berries contain different types of polyphenols including flavonoids and phenolic acids. Anthocyanin is the major type of flavonoid. C3G is the most prominent type of anthocyanin, which accounts for 79% of total anthocyanin in all extracts. The variety Larissa at H5 contained the highest average C3G content, and its ethanol extract had the highest (1212.3±63.9 mg/100g FW) while, Evie at H1 contained the lowest C3G content (96.9±40.4 mg/100g FW). The average C3G content of Larissa from H1 – H5 varies from 208 – 1212 mg/100g FW. Quarcetin-3-Rutinoside (Q3Rut) is the major type of flavonol and highest is observed in Rebecca at H4 (47.81 mg/100g FW). The haskap berries also contained phenolic acids, but approximately 95% of the phenolic acids consisted of chlorogenic acid. The cultivar Larissa has a higher level of anthocyanin than the other four cultivars. The highest total phenolic content is observed in Evie at H5 (2.97±1.03 mg/g DW) while the lowest in Rebecca at H1 (1.47±0.96 mg/g DW). The antioxidant capacity of Evie at H5 was higher (14.40±2.21 µmol TE/ g DW) among other cultivars and the lowest observed in Aurora at H3 (5.69±0.34 µmol TE/ g DW). Furthermore, Larissa H5 shows the greatest inhibition of carbohydrate-hydrolyzing enzymes including alpha-glucosidase and alpha-amylase. In conclusion Larissa, at H5 demonstrated highest polyphenol composition and antidiabetic properties.

Keywords: anthocyanin, cyanidin-3-O-glucoside, haskap, type 2 diabetes

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

Abstract:

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 244

Authors: P. Chaijak, M. Lertworapreecha, C. Sukkasem

Abstract:

Extracellular laccases are copper-containing microbial enzymes with many industrial biotechnological applications. This study evaluated the ability of nutrients in coconut coir to enhance the yield of extracellular laccase of Galactomyces reesii IFO 10823 and develop a co-culture between this yeast and other filamentous fungi isolated from the fungus comb of Macrotermes sp. The co-culture between G. reesii IFO 10823 and M. indicus FJ-M-5 (G3) gave the highest activity at 580.20 U/mL. When grown in fermentation media prepared from coconut coir and distilled water at 70% of initial moisture without supplement addition, G3 produced extracellular laccase of 113.99 U/mL.

Keywords: extracellular laccase, production, yeast, natural inducer

Procedia PDF Downloads 252

Authors: Sesethu G. Njokweni, Marelize Botes, Emile W. H. Van Zyl

Abstract:

An alternative strategy to the production of bioethanol is by examining the degradability of biomass in a natural system such as the rumen of mammals. This anaerobic microbial community has higher cellulolytic activities than microbial communities from other habitats and degrades cellulose to produce volatile fatty acids (VFA), methane and CO₂. VFAs have the potential to serve as intermediate products for electrochemical conversion to hydrocarbon fuels. In vitro mimicking of this process would be more cost-effective than bioethanol production as it does not require chemical pre-treatment of biomass, a sterile environment or added enzymes. The strategies of the carboxylate platform and the co-cultures of a bovine ruminal microbiota from cannulated cows were combined in order to investigate and optimize the bioconversion of agricultural biomass (apple and grape pomace, citrus pulp, sugarcane bagasse and triticale straw) to high value VFAs as intermediates for biofuel production in a consolidated bioprocess. Optimisation of reactor conditions was investigated using five different ruminal inoculum concentrations; 5,10,15,20 and 25% with fixed pH at 6.8 and temperature at 39 ˚C. The ANKOM 200/220 fiber analyser was used to analyse in vitro neutral detergent fiber (NDF) disappearance of the feedstuffs. Fresh and cryo-frozen (5% DMSO and 50% glycerol for 3 months) rumen cultures were tested for the retainment of fermentation capacity and durability in 72 h fermentations in 125 ml serum vials using a FURO medical solutions 6-valve gas manifold to induce anaerobic conditions. Fermentation of apple pomace, triticale straw, and grape pomace showed no significant difference (P > 0.05) in the effect of 15 and 20 % inoculum concentrations for the total VFA yield. However, high performance liquid chromatographic separation within the two inoculum concentrations showed a significant difference (P < 0.05) in acetic acid yield, with 20% inoculum concentration being the optimum at 4.67 g/l. NDF disappearance of 85% in 96 h and total VFA yield of 11.5 g/l in 72 h (A/P ratio = 2.04) for apple pomace entailed that it was the optimal feedstuff for this process. The NDF disappearance and VFA yield of DMSO (82% NDF disappearance and 10.6 g/l VFA) and glycerol (90% NDF disappearance and 11.6 g/l VFA) stored rumen also showed significantly similar degradability of apple pomace with lack of treatment effect differences compared to a fresh rumen control (P > 0.05). The lack of treatment effects was a positive sign in indicating that there was no difference between the stored samples and the fresh rumen control. Retaining of the fermentation capacity within the preserved cultures suggests that its metabolic characteristics were preserved due to resilience and redundancy of the rumen culture. The amount of degradability and VFA yield within a short span was similar to other carboxylate platforms that have longer run times. This study shows that by virtue of faster rates and high extent of degradability, small scale alternatives to bioethanol such as rumen microbiomes and other natural fermenting microbiomes can be employed to enhance the feasibility of biofuels large-scale implementation.

Keywords: agricultural wastes, carboxylate platform, rumen microbiome, volatile fatty acids

Procedia PDF Downloads 117

Authors: Zoi Konsoula

Abstract:

The increasing prevalence of Alzheimer’s disease (AD) and diabetes mellitus (DM) constitutes them major global health problems. Recently, the inhibition of key enzyme activity is considered a potential treatment of both diseases. Specifically, inhibition of acetylcholinesterase (AChE), the key enzyme involved in the breakdown of the neurotransmitter acetylcholine, is a promising approach for the treatment of AD, while inhibition of α-amylase retards the hydrolysis of carbohydrates and, thus, reduces hyperglycemia. Unfortunately, commercially available AChE and α-amylase inhibitors are reported to possess side effects. Consequently, there is a need to develop safe and effective treatments for both diseases. In the present study, pomegranate peel (PP) was extracted using various solvents of increasing polarity, while two extraction methods were employed, the conventional maceration and the ultrasound assisted extraction (UAE). The concentration of bioactive phytoconstituents, such as total phenolics (TPC) and total flavonoids (TFC) in the prepared extracts was evaluated by the Folin-Ciocalteu and the aluminum-flavonoid complex method, respectively. Furthermore, the anti-neurodegenerative and anti-hyperglycemic activity of all extracts was determined using AChE and α-amylase inhibitory activity assays, respectively. The inhibitory activity of the extracts against AChE and α-amylase was characterized by estimating their IC₅₀ value using a dose-response curve, while galanthamine and acarbose were used as positive controls, respectively. Finally, the kinetics of AChE and α-amylase in the presence of the most inhibitory potent extracts was determined by the Lineweaver-Burk plot. The methanolic extract prepared using the UAE contained the highest amount of phytoconstituents, followed by the respective ethanolic extract. All extracts inhibited acetylcholinesterase in a dose-dependent manner, while the increased anticholinesterase activity of the methanolic (IC₅₀ = 32 μg/mL) and ethanolic (IC₅₀ = 42 μg/mL) extract was positively correlated with their TPC content. Furthermore, the activity of the aforementioned extracts was comparable to galanthamine. Similar results were obtained in the case of α-amylase, however, all extracts showed lower inhibitory effect on the carbohydrate hydrolyzing enzyme than on AChE, since the IC₅₀ value ranged from 84 to 100 μg/mL. Also, the α-amylase inhibitory effect of the extracts was lower than acarbose. Finally, the methanolic and ethanolic extracts prepared by UAE inhibited both enzymes in a mixed (competitive/noncompetitive) manner since the Kₘ value of both enzymes increased in the presence of extracts, while the Vmax value decreased. The results of the present study indicate that PP may be a useful source of active compounds for the management of AD and DM. Moreover, taking into consideration that PP is an agro-industrial waste product, its valorization could not only result in economic efficiency but also reduce the environmental pollution.

Keywords: acetylcholinesterase, Alzheimer’s disease, α-amylase, diabetes mellitus, pomegranate

Procedia PDF Downloads 111

Authors: Fadila Mohamed Mahmoud., Derkaoui I., Krimi Z.

Abstract:

Date palm is a plant which presents a very good adaptation to the difficult conditions of the environment in particular to the drought and saline stress even at high temperatures. This adaptation is related on the biology of the plant and to the presence of a microflora endophyte which live inside its tissues. Fifteen endophytics fungi isolated from date palm were tested in vitro in the presence of various NaCl concentrations to select halotolerantes isolates. These same endophytes were tested for their colonizing capacity by the description of the production of secondary metabolites more particularly the enzymes (pectinases, proteases, and phosphorylases), and the production of antibiotics and growth hormones. Significant difference was observed between the isolates with respect to the tests carried out.

Keywords: Date palm, Halotolerantes, endophyte, Secondary metabolites.

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Authors: Fuad Ameen, Mohamed Moslem, Sarfaraz Hadi

Abstract:

Twenty-eight fungal isolates belonging to 12 genera were derived from debris, sediment and water samples collected from Avicennia marina stands 25km south of Jeddah city on the Red Sea coast of Saudi Arabia. Eight of these isolates were found to be able to grow in association cellulosic waste materials under in vitro conditions in the absence of any carbon source. Isolates were further tested for their potential to degrade paper and clothes wastes by co-cultivation under aeration on a rotary shaker. These fungi accumulated significantly higher biomass, produced ligninolytic and cellulase enzymes, and liberated larger volumes of CO2. These observations indicated that the selected isolates were able to break down and consume the waste materials.

Keywords: biodegradation, enzyme activity, waste materials, mangrove

Procedia PDF Downloads 544

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

Abstract:

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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