Search results for: liver toxicity
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1626

Search results for: liver toxicity

1146 Development of Electrochemical Biosensor Based on Dendrimer-Magnetic Nanoparticles for Detection of Alpha-Fetoprotein

Authors: Priyal Chikhaliwala, Sudeshna Chandra

Abstract:

Liver cancer is one of the most common malignant tumors with poor prognosis. This is because liver cancer does not exhibit any symptoms in early stage of disease. Increased serum level of AFP is clinically considered as a diagnostic marker for liver malignancy. The present diagnostic modalities include various types of immunoassays, radiological studies, and biopsy. However, these tests undergo slow response times, require significant sample volumes, achieve limited sensitivity and ultimately become expensive and burdensome to patients. Considering all these aspects, electrochemical biosensors based on dendrimer-magnetic nanoparticles (MNPs) was designed. Dendrimers are novel nano-sized, three-dimensional molecules with monodispersed structures. Poly-amidoamine (PAMAM) dendrimers with eight –NH₂ groups using ethylenediamine as a core molecule were synthesized using Michael addition reaction. Dendrimers provide added the advantage of not only stabilizing Fe₃O₄ NPs but also displays capability of performing multiple electron redox events and binding multiple biological ligands to its dendritic end-surface. Fe₃O₄ NPs due to its superparamagnetic behavior can be exploited for magneto-separation process. Fe₃O₄ NPs were stabilized with PAMAM dendrimer by in situ co-precipitation method. The surface coating was examined by FT-IR, XRD, VSM, and TGA analysis. Electrochemical behavior and kinetic studies were evaluated using CV which revealed that the dendrimer-Fe₃O₄ NPs can be looked upon as electrochemically active materials. Electrochemical immunosensor was designed by immobilizing anti-AFP onto dendrimer-MNPs by gluteraldehyde conjugation reaction. The bioconjugates were then incubated with AFP antigen. The immunosensor was characterized electrochemically indicating successful immuno-binding events. The binding events were also further studied using magnetic particle imaging (MPI) which is a novel imaging modality in which Fe₃O₄ NPs are used as tracer molecules with positive contrast. Multicolor MPI was able to clearly localize AFP antigen and antibody and its binding successfully. Results demonstrate immense potential in terms of biosensing and enabling MPI of AFP in clinical diagnosis.

Keywords: alpha-fetoprotein, dendrimers, electrochemical biosensors, magnetic nanoparticles

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1145 Cedrela Toona Roxb.: An Exploratory Study Describing Its Antidiabetic Property

Authors: Kinjal H. Shah, Piyush M. Patel

Abstract:

Diabetes mellitus is considered to be a serious endocrine syndrome. Synthetic hypoglycemic agents can produce serious side effects including hematological effects, coma, and disturbances of the liver and kidney. In addition, they are not suitable for use during pregnancy. In recent years, there have been relatively few reports of short-term side effects or toxicity due to sulphonylureas. Published figures and frequency of side effects in large series of patient range from about 1 to 5%, with symptoms severe enough to lead to the withdrawal of the drug in less than 1 to 2%. Adverse effects, in general, have been of the following type: allergic skin reactions, gastrointestinal disturbances, blood dyscrasias, hepatic dysfunction, and hypoglycemia. The associated disadvantages with insulin and oral hypoglycemic agents have led to stimulation in the research for locating natural resources showing antidiabetic activity and to explore the possibilities of using traditional medicines with proper chemical and pharmacological profiles. Literature survey reveals that the inhabitants of Abbottabad district of Pakistan use the dried leaf powder along with table salt and water orally for treating diabetes, skin allergy, wounds and as a blood purifier, where they pronounced the plant locally as ‘Nem.' The detailed phytochemical investigation of the Cedrela toona Roxb. leaves for antidiabetic activity has not been documented. Hence, there is a need for phytochemical investigation of the leaves for antidiabetic activity. The collection of fresh leaves and authentification followed by successive extraction, phytochemical screening, and testing of antidiabetic activity. The blood glucose level was reduced maximum in ethanol extract at 5th and 7th h after treatment. Blood glucose was depressed by 8.2% and 10.06% in alloxan – induced diabetic rats after treatment which was comparable to the standard drug, Glibenclamide. This may be due to the activation of the existing pancreatic cells in diabetic rats by the ethanolic extract.

Keywords: antidiabetic, Cedrela toona Roxb., phytochemical screening, blood glucose

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1144 In vitro Investigation of Genotoxic and Antigenotoxic Properties of Gunnera perpensa Roots Extracts

Authors: P. H. Mfengwana, S. S. Mashele, L. Verschaeve, R. Anthonissen, I. T. Manduna

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Gunnera perpensa is traditionally used mostly by women for the treatment of different gynaecological related conditions due to its proven uterine contractility effects. The uses of this plant include menstrual pain relief, treatment of infertility and promotion of easy labour. However, even though this plant species has been reported to possess numerous medicinal properties, to author’s best knowledge, its safety has not been investigated. Thus, this study was aimed at investigating the genotoxicity and antigenotoxicity of Gunnera perpensa aqueous, methanol and dichloromethane extracts. The in vitro toxicity of the plant extracts was assessed with the neutral red uptake (NRU) test. Genotoxic and antigenotoxic properties of Gunnera perpensa were investigated using high-throughput assays: bacterial Vitotox test and the alkaline comet assay with and without S9 activation on human C3A cells. Ethyl Methanesulfonate (EMS) and 4-nitroquinoline-oxide (4-NQO) were used as positive controls, respectively. All extracts showed toxicity in a dose-dependent manner; however, that does not mean they were all genotoxic. Methanol extract did show genotoxicity with S9 (metabolism) only at the highest concentration of 500 µg/ml due to increased DNA damage observed, however, no genotoxicity was observed from other concentrations. Therefore, the results show that Gunnera perpensa extracts are genotoxic and not safe for human use.

Keywords: antigenotoxicity, comet test, genotoxicity, Gunnera perpensa, vitotox assay

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1143 Bone Mineralization in Children with Wilson’s Disease

Authors: Shiamaa Eltantawy, Gihan Sobhy, Alif Alaam

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Wilson disease, or hepatolenticular degeneration, is an autosomal recessive disease that results in excess copper buildup in the body. It primarily affects the liver and basal ganglia of the brain, but it can affect other organ systems. Musculoskeletal abnormalities, including premature osteoarthritis, skeletal deformity, and pathological bone fractures, can occasionally be found in WD patients with a hepatic or neurologic type. The aim was to assess the prevalence of osteoporosis and osteopenia in Wilson’s disease patients. This case-control study was conducted on ninety children recruited from the inpatient ward and outpatient clinic of the Paediatric Hepatology, Gastroenterology, and Nutrition department of the National Liver Institute at Menofia University, aged from 1 to 18 years. Males were 49, and females were 41. Children were divided into three groups: (Group I) consisted of thirty patients with WD; (Group II) consisted of thirty patients with chronic liver disease other than WD; (Group III) consisted of thirty age- and sex-matched healthy The exclusion criteria were patients with hyperparathyroidism, hyperthyroidism, renal failure, Cushing's syndrome, and patients on certain drugs such as chemotherapy, anticonvulsants, or steroids. All patients were subjected to the following: 1- Full history-taking and clinical examination. 2-Laboratory investigations: (FBC,ALT,AST,serum albumin, total protein, total serum bilirubin,direct bilirubin,alkaline phosphatase, prothrombin time, serum critine,parathyroid hormone, serum calcium, serum phosphrus). 3-Bone mineral density (BMD, gm/cm2) values were measured by dual-energy X-ray absorptiometry (DEXA). The results revealed that there was a highly statistically significant difference between the three groups regarding the DEXA scan, and there was no statistically significant difference between groups I and II, but the WD group had the lowest bone mineral density. The WD group had a large number of cases of osteopenia and osteoporosis, but there was no statistically significant difference with the group II mean, while a high statistically significant difference was found when compared to group III. In the WD group, there were 20 patients with osteopenia, 4 patients with osteoporosis, and 6 patients who were normal. The percentages were 66.7%, 13.3%, and 20%, respectively. Therefore, the largest number of cases in the WD group had osteopenia. There was no statistically significant difference found between WD patients on different treatment regimens regarding DEXA scan results (Z-Score). There was no statistically significant difference found between patients in the WD group (normal, osteopenic, or osteoporotic) regarding phosphorus (mg/dL), but there was a highly statistically significant difference found between them regarding ionised Ca (mmol/L). Therefore, there was a decrease in bone mineral density when the Ca level was decreased. In summary, Wilson disease is associated with bone demineralization. The largest number of cases in the WD group in our study had osteopenia (66.7%). Different treatment regimens (zinc monotherapy, Artamin, and zinc) as well as different laboratory parameters have no effect on bone mineralization in WD cases. Decreased ionised Ca is associated with low BMD in WD patients. Children with WD should be investigated for BMD.

Keywords: wilson disease, Bone mineral density, liver disease, osteoporosis

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1142 Chronic Renal Failure Associated with Heavy Metal Contamination of Drinking Water in Hail, Kingdom of Saudi Arabia

Authors: Elsayed A. M. Shokr, A. Alhazemi, T. Naser, Talal A. Zuhair, Adel A. Zuhair, Ahmed N. Alshamary, Thamer A. Alanazi, Hosam A. Alanazi

Abstract:

The main threats to human health from heavy metals are associated with exposure to Pb, Cd, Cu, Mo, Zn, Ni, Mn Co and Cr. is mainly via intake of drinking water being the most important source in most populations. These metals have been extensively studied and their effects on human health regularly reviewed by international bodies such as the WHO. Heavy metals have been used by humans for thousands of years. Although several adverse health effects of heavy metals have been known for a long time, exposure to heavy metals continues, and is even increasing in some parts of the world, in particular in less developed countries, though emissions have declined in most developed countries over the last 100 years. A strong relationship between contaminated drinking water with heavy metals from some of the stations of water shopping in Hail, KSA and chronic diseases such as renal failure, liver cirrhosis, and chronic anemia has been identified in this study. These diseases are apparently related to contaminant drinking water with heavy metals such as Pb, Cd, Cu, Mo, Zn, Ni, Mn Co and Cr. Renal failure is related to contaminate drinking water with lead and cadmium, liver cirrhosis to copper and molybdenum, and chronic anemia to copper and cadmium. Recent data indicate that adverse health effects of cadmium exposure may occur at lower exposure levels than previously anticipated, primarily in the form of kidney damage but possibly also bone effects and fractures. The general population is primarily exposed to mercury via drinking water being a major source of methyl mercury exposure, and dental amalgam. During the last century lead, cadmium, zinc, iron and arsenic is mainly via intake of drinking water being the most important source in most populations. Long-term exposure to lead, cadmium, zinc, iron and arsenic in drinking-water is mainly related to primarily in the form of kidney damage. Studies of these diseases suggest that abnormal incidence in specific areas is related to toxic materials in the groundwater and thereby led to the contamination of drinking water in these areas.

Keywords: heavy metals, liver functions, kidney functions and chronic renal failure, hail, renal, water

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1141 Caecotrophy Behaviour of the Rabbits (Oryctolagus cuniculus)

Authors: Awadhesh Kishore

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One of the most unique characteristics of rabbit feeding behaviour is caecotrophy, which involves the excretion and immediate consumption of specific faeces known as soft faeces. Caecotrophy in rabbits is the instinctual behaviour of eating soft faeces; reduced caecotrophy decreases rabbit growth and lipid synthesis in the liver. Caecotroph ingestion is highest when rabbits are fed a diet high in indigestible fibre. The colon produces two types of waste: hard and soft pellets. The hard pellets are expelled, but the soft pellets are re-ingested by the rabbit directly upon being expelled from the anus by twisting itself around and sucking in those pellets as they emerge from the anus. The type of alfalfa hay in the feed of the rabbits does not affect volatile fatty acid concentration, the pattern of fermentation, or pH in the faeces. The cecal content and the soft faeces contain significant amounts of retinoids and carotenoids, while in the tissues (blood, liver, and kidney), these pigments do not occur in substantial amounts. Preventing caecotrophy reduced growth and altered lipid metabolism, depressing the development of new approaches for rabbit feeding and production. Relative abundance is depressed for genes related to metabolic pathways such as vitamin C and sugar metabolism, vitamin B2 metabolism, and bile secretion. The key microorganisms that regulate the rapid growth performance of rabbits may provide useful references for future research and the development of microecological preparations.

Keywords: caecocolonic microorganisms, caecotrophy, fasting caecotrophy, rabbits, soft pellets

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1140 Ultrastructural Characterization of Lipid Droplets of Rat Hepatocytes after Whole Body 60-Cobalt Gamma Radiation

Authors: Ivna Mororó, Lise P. Labéjof, Stephanie Ribeiro, Kely Almeida

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Lipid droplets (LDs) are normally presented in greater or lesser number in the cytoplasm of almost all eukaryotic and some prokaryotic cells. They are independent organelles composed of a lipid ester core and a surface phospholipid monolayer. As a lipid storage form, they provide an available source of energy for the cell. Recently it was demonstrated that they play an important role in other many cellular processes. Among the many unresolved questions about them, it is not even known how LDs is formed, how lipids are recruited to LDs and how they interact with the other organelles. Excess fat in the organism is pathological and often associated with the development of some genetic, hormonal or behavioral diseases. The formation and accumulation of lipid droplets in the cytoplasm can be increased by exogenous physical or chemical agents. It is well known that ionizing radiation affects lipid metabolism resulting in increased lipogenesis in cells, but the details of this process are unknown. To better understand the mode of formation of LDs in liver cells, we investigate their ultrastructural morphology after irradiation. For that, Wistar rats were exposed to whole body gamma radiation from 60-cobalt at various single doses. Samples of the livers were processed for analysis under a conventional transmission electron microscope. We found that when compared to controls, morphological changes in liver cells were evident at the higher doses of radiation used. It was detected a great number of lipid droplets of different sizes and homogeneous content and some of them merged each other. In some cells, it was observed diffused LDs, not limited by a monolayer of phospholipids. This finding suggests that the phospholipid monolayer of the LDs was disrupted by ionizing radiation exposure that promotes lipid peroxydation of endo membranes. Thus the absence of the phospholipid monolayer may prevent the realization of some cellular activities as follow: - lipid exocytosis which requires the merging of LDs membrane with the plasma membrane; - the interaction of LDs with other membrane-bound organelles such as the endoplasmic reticulum (ER), the golgi and mitochondria and; - lipolysis of lipid esters contained in the LDs which requires the presence of enzymes located in membrane-bound organelles as ER. All these impediments can contribute to lipid accumulation in the cytoplasm and the development of diseases such as liver steatosis, cirrhosis and cancer.

Keywords: radiobiology, hepatocytes, lipid metabolism, transmission electron microscopy

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1139 Theocritus’ Idylls and the Serenading of Mystical Women: Toxic Modes of Seduction in Pastoral Literature

Authors: Kayla Fanning

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Theocritus’ use of near-pastoral motifs in creating the lamenting narrative in “The Sorceress” idyll suggests a link between Simaetha and the quintessential shepherd that ultimately transcends the bucolic serenading structure, evident in “The Serenade”, as it pertains to depictions of women. In Theocritus’ “The Serenade”, an anonymous goatherd serenades his beloved, Amaryllis, in hopes of persuading her to reciprocate his love. This serenade soon turns into a vicious lament where all hope for reciprocation dissolves, leaving the goatherd severely melancholic and malignant. In “The Cyclops’ Serenade”, the cyclops, Polyphemus, sings of Galatea in solitude; in so doing, he negotiates between feelings of heartache and anger that eventually subside. His depiction of Galatea, in being less vindictive than the goatherd’s, manifests less toxicity. In adopting, and essentially creating, this serenading structure, Theocritus illustrates his ability to alter portrayals of women while maintaining the premise of the pastoral serenade; that is the shepherd's lament of his indifferent beloved. A thematic intertextual analysis of the idylls reveals a variety of ways in which the toxicity of the goatherd’s relation to Amaryllis is mutated or even inverted. In “The Sorceress”, a powerful witch named Simaetha spellbinds her unfaithful lover and angrily laments his betrayal in a way that is reminiscent of the goatherd's harmful behavior towards Amaryllis.

Keywords: femininity, pastoral, serenade, Theocritus

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1138 A Comparative Approach for Modeling the Toxicity of Metal Mixtures in Two Ecologically Related Three-Spined (Gasterosteus aculeatus L.) And Nine-Spined (Pungitius pungitius L.) Sticklebacks

Authors: Tomas Makaras

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Sticklebacks (Gasterosteiformes) are increasingly used in ecological and evolutionary research and become well-established role as model species for biologists. However, ecotoxicology studies concerning behavioural effects in sticklebacks regarding stress responses, mainly induced by chemical mixtures, have hardly been addressed. Moreover, although many authors in their studies emphasised the similarity between three-spined and nine-spined stickleback in morphological, neuroanatomical and behavioural adaptations to environmental changes, several comparative studies have revealed considerable differences between these species in and their susceptibility and resistance to variousstressors in laboratory experiments. The hypothesis of this study was that three-spined and nine-spined stickleback species will demonstrate apparent differences in response patterns and sensitivity to metal-based chemicals stimuli. For this purpose, we investigated the swimming behaviour (including mortality rate based on 96-h LC50 values) of two ecologically similar three-spined (Gasterosteusaculeatus) and nine-spined sticklebacks (Pungitiuspungitius) to short-term (up to 24 h) metal mixture (MIX) exposure. We evaluated the relevance and efficacy of behavioural responses of test species in the early toxicity assessment of chemical mixtures. Fish exposed to six (Zn, Pb, Cd, Cu, Ni and Cr) metals in the mixture were either singled out by the Water Framework Directive as priority or as relevant substances in surface water, which was prepared according to the environmental quality standards (EQSs) of these metals set for inland waters in the European Union (EU) (Directive 2013/39/EU). Based on acute toxicity results, G. aculeatus found to be slightly (1.4-fold) more tolerant of MIX impact than those of P. pungitius specimens. The performed behavioural analysis showed the main effect on the interaction between time, species and treatment variables. Although both species exposed to MIX revealed a decreasing tendency in swimming activity, these species’ responsiveness to MIX was somewhat different. Substantial changes in the activity of G. aculeatus were established after 3-h exposure to MIX solutions, which was 1.43-fold lower, while in the case of P. pungitius, 1.96-fold higher than established 96-h LC50 values for each species. This study demonstrated species-specific differences in response sensitivity to metal-based water pollution, indicating behavioural insensitivity of P. pungitiuscompared to G. aculeatus. While many studies highlight the usefulness and suitability of nine-spined sticklebacks for evolutionary and ecological research, attested by their increasing popularity in these fields, great caution must be exercised when using them as model species in ecotoxicological research to probe metal contamination. Meanwhile, G. aculeatus showed to be a promising bioindicator species in the environmental ecotoxicology field.

Keywords: acute toxicity, comparative behaviour, metal mixture, swimming activity

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1137 Enzymatic Biomonitoring of Aquatic Pollution at Jeddah Southern Red Sea Shore

Authors: Saleh Mohamed, Mohamed El-Shal, Taha Kumosani, Ahmad Mal, Youssri Ahmed, Yasser Almulaiky

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The marine environment of the Jeddah southern red sea shore is subjected to increasing anthropogenic activities as sewage sludge draining and desalting processes. The objective of this study is to compare the quantitative responses of enzymatic biomarkers in fish from polluted area with the responses of organism from reference area. Enzymatic biomarkers as neurotoxic, antioxidant and detoxifying enzymes were evaluated in the brain and liver from Variola louti as a sentinel species sampled from both polluted and reference sites in the Jeddah southern red sea shore during four months January, April, July and October in 2014 and 2015. In brain of V. louti, the activity of acetylcholinestease (AChE) collected from reference area significantly increased 8.8 and 10.5 folds than that from polluted area in 2014 and 2015, respectively. The activities of catalase (CAT), glutathione reductase (GR) and glutathione peroxidase (GPx) and glutathione-S-transferase (GST) from liver of V. louti in polluted area significantly increased 1.4, 1.27 and 3, 4.5 and 4.37, 2 and 5, 4.5 folds than that from reference area in 2014 and 2015, respectively. The levels of examined enzymes are approximately similar in the four seasons detected in 2014 and 2015 indicating that the similar components of sewage were draining in red sea. In conclusion, these findings suggest the important of enzymatic biomarkers in monitoring the pollution in Jeddah red sea shore.

Keywords: Variola louti, enzymatic biomarkers, pollution, Red sea

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1136 Development and Characterization of Topical 5-Fluorouracil Solid Lipid Nanoparticles for the Effective Treatment of Non-Melanoma Skin Cancer

Authors: Sudhir Kumar, V. R. Sinha

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Background: The topical and systemic toxicity associated with present nonmelanoma skin cancer (NMSC) treatment therapy using 5-Fluorouracil (5-FU) make it necessary to develop a novel delivery system having lesser toxicity and better control over drug release. Solid lipid nanoparticles offer many advantages like: controlled and localized release of entrapped actives, nontoxicity, and better tolerance. Aim:-To investigate safety and efficacy of 5-FU loaded solid lipid nanoparticles as a topical delivery system for the treatment of nonmelanoma skin cancer. Method: Topical solid lipid nanoparticles of 5-FU were prepared using Compritol 888 ATO (Glyceryl behenate) as lipid component and pluronic F68 (Poloxamer 188), Tween 80 (Polysorbate 80), Tyloxapol (4-(1,1,3,3-Tetramethylbutyl) phenol polymer with formaldehyde and oxirane) as surfactants. The SLNs were prepared with emulsification method. Different formulation parameters viz. type and ratio of surfactant, ratio of lipid and ratio of surfactant:lipid were investigated on particle size and drug entrapment efficiency. Results: Characterization of SLNs like–Transmission Electron Microscopy (TEM), Differential Scannig calorimetry (DSC), Fourier transform infrared spectroscopy (FTIR), Particle size determination, Polydispersity index, Entrapment efficiency, Drug loading, ex vivo skin permeation and skin retention studies, skin irritation and histopathology studies were performed. TEM results showed that shape of SLNs was spherical with size range 200-500nm. Higher encapsulation efficiency was obtained for batches having higher concentration of surfactant and lipid. It was found maximum 64.3% for SLN-6 batch with size of 400.1±9.22 nm and PDI 0.221±0.031. Optimized SLN batches and marketed 5-FU cream were compared for flux across rat skin and skin drug retention. The lesser flux and higher skin retention was obtained for SLN formulation in comparison to topical 5-FU cream, which ensures less systemic toxicity and better control of drug release across skin. Chronic skin irritation studies lacks serious erythema or inflammation and histopathology studies showed no significant change in physiology of epidermal layers of rat skin. So, these studies suggest that the optimized SLN formulation is efficient then marketed cream and safer for long term NMSC treatment regimens. Conclusion: Topical and systemic toxicity associated with long-term use of 5-FU, in the treatment of NMSC, can be minimized with its controlled release with significant drug retention with minimal flux across skin. The study may provide a better alternate for effective NMSC treatment.

Keywords: 5-FU, topical formulation, solid lipid nanoparticles, non melanoma skin cancer

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1135 Evaluation of Toxicity of Cerium Oxide on Zebrafish Developmental Stages

Authors: Roberta Pecoraro, Elena Maria Scalisi

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Engineered Nanoparticles (ENPs) and Nanomaterials (ENMs) concern an active research area and a sector in full expansion. They have physical-chemical characteristics and small size that improve their performance compared to common materials. Due to the increase in their production and their subsequent release into the environment, new strategies are emerging to assess risk of nanomaterials. NPs can be released into the environment through aquatic systems by human activities and exert toxicity on living organisms. We evaluated the potential toxic effect of cerium oxide (CeO2) nanoparticles because it’s used in different fields due to its peculiar properties. In order to assess nanoparticles toxicity, Fish Embryo Toxicity (FET) test was performed. Powders of CeO2 NPs supplied by the CNR-IMM of Catania are indicated as CeO2 type 1 (as-prepared) and CeO2 type 2 (modified), while CeO2 type 3 (commercial) is supplied by Sigma-Aldrich. Starting from a stock solution (0.001g/10 ml dilution water) of each type of CeO2 NPs, the other concentration solutions were obtained adding 1 ml of the stock solution to 9 ml of dilution water, leading to three different solutions of concentration (10-4, 10-5, 10-6 g/ml). All the solutions have been sonicated to avoid natural tendency of NPs to aggregate and sediment. FET test was performed according to the OECD guidelines for testing chemicals using our internal protocol procedure. A number of eight selected fertilized eggs were placed in each becher filled with 5 ml of each concentration of the three types of CeO2 NPs; control samples were incubated only with dilution water. Replication was performed for each concentration. During the exposure period, we observed four endpoints (embryo coagulation, lack of formation of somites, failure to lift the yolk bag, no heartbeat) by a stereomicroscope every 24 hours. Immunohistochemical analysis on treated larvae was performed to evaluate the expression of metallothioneins (MTs), Heat Shock Proteins 70 (HSP70) and 7-ethoxyresorufin-O-diethylase (EROD). Our results have not shown evident alterations on embryonic development because all embryos completed the development and the hatching of the eggs, started around the 48th hour after exposure, took place within the last observation at 72 hours. A good reactivity, both in the embryos and in the newly hatched larvae, was found. The presence of heartbeat has also been observed in embryos with reduced mobility confirming their viability. A higher expression of EROD biomarker was observed in the larvae exposed to the three types of CeO2, showing a clear difference with the control. A weak positivity was found for MTs biomarker in treated larvae as well as in the control. HSP70 are expressed homogeneously in all the type of nanoparticles tested but not too much greater than control. Our results are in agreement with other studies in the literature, in which the exposure of Danio rerio larvae to other metal oxide nanoparticles does not show adverse effects on survival and hatching time. Further studies are necessary to clarify the role of these NPs and also to solve conflicting opinions.

Keywords: Danio rerio, endpoints, fish embryo toxicity test, metallic nanoparticles

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1134 Gallbladder Amyloidosis Causing Gangrenous Cholecystitis: A Case Report

Authors: Christopher Leung, Guillermo Becerril-Martinez

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Amyloidosis is a rare systemic disease where abnormal proteins invade various organs and impede their function. Occasionally, they can manifest in a solidary organ such as the heart, lung, and nervous systems; rarely do they manifest in the gallbladder. Diagnosis often requires biopsy of the affected area and histopathology shows deposition of abnormally folded globular proteins called amyloid proteins. This case presents a 69-year-old male with a 3-month history of RUQ pain, diarrhea and non-specific symptoms of tiredness, etc. On imaging, both his US and CT abdomen showed gallbladder wall thickening and pericholecystic fluid, which may represent acute cholecystitis with hypodense lesions around the gallbladder, possibly representing liver abscesses. Given his symptoms of abdominal pain and imaging findings, this gentleman eventually had a laparoscopic cholecystectomy showing a gangrenous gallbladder with a mass on the liver bed. On histopathology, it showed amorphous hyaline eosinophilic material, which Congo-stained confirmed amyloidosis. Amyloidosis explained his non-specific symptoms, he avoided further biopsy, and he was commenced immediately on Lenalidomide. Involvement of the gallbladder is extremely rare, with less than 30 cases around the world. Half of the cases are reported as primary amyloidosis. This case adds to the current literature regarding primary gallbladder amyloidosis. Importantly, this case highlights how laparoscopic cholecystectomy can help with the diagnosis of gallbladder amyloidosis.

Keywords: amyloidosis, cholecystitis, gangrenous cholecystitis, gallbladder, systemic amyloidosis

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1133 Pathological and Molecular Diagnosis of Caseous Lymphadenitis in Chinkara Deer (Gazella Bennettii), in Pakistan

Authors: Mudassar Iqbal, Riaz Hussain, Khalid Mehmood, Farah Ali, Fazal Mahmood, Abdul Ghaffar

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Corynebacterium pseudotuberculosis is an important cause of caseous lymphadenitis (CL), a complex, chronic devastating and destructive disease of small ruminants. In present study, postmortem examination of Chinkara deer (n=25) was conducted in year 2014. Pus samples suggestive of CL were collected from the superficial lymph nodes, liver, spleen and lungs during necropsy and subjected to standard microbiological procedures for isolation and molecular analysis of bacterial pathogens. Pus samples collected from carcasses (25) presenting clinical lesions of C. pseudotuberculosis infection was identified in 19 (76%) carcasses on the basis of culture characteristics. The frequency of C. pseudotuberculosis bacterium was higher in older animals as compared to young animals. Grossly, multiple tubercles of variable size having caseous material were observed in liver, lungs, spleen and lymph nodes. Histopathologically, tissue sections from all the visceral organs were extensively plugged with abscess. In present study specific prolineiminopeptidase (PIP) gene of the C. pseudotuberculosis was amplified by the Polymerase chain reaction technique (PCR) in 17(25) cases. The efficient and reliable molecular analysis along with necropsy findings in present study can be used as valuable approach for diagnosis of caseous lymphadenitis in small ruminants.

Keywords: Chinkara deer, Corynebacterium pseudotuberculosis, Caseous lymphadenitis, PCR

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1132 Toxicity of Biopesticide Metarhizium anisopliae var acridium "Green Muscle" on the Cuticle of the Desert Locust Schistocerca gegaria (Forskål, 1775)

Authors: F. Haddadj, F. Acheuk, S. Hamdi, S. Zenia, A. Smai, H. Saadi, B. Doumandji-Mitiche

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Locust is causing significant losses in agricultural production in the countries concerned by the invasion. Up to the present control strategy has consisted only of the spreaders chemicals; they have proven harmful to the environment and. For this, a new control method appeared it comes to the biological control based mostly by using microorganism. It is in that sense is we've made our contribution by the use of a biopesticide which is entomopathogenic fungus Metarhizium anisopliae var acridium "Green Muscle" on part of the cuticule the larval of fifth instar locust Schistocerca gregaria (Forskål, 1775). Preliminary test on the study of the pathogenicity of M. anisopliae var acridium biocontrol agent, was conducted in the laboratory on L5 S. gregaria, on which we inoculated treatment in the digestive tract and it administrant 20μl of entomopathogenic solution orally at a dose DL50 = 3.25 x107 sp./ ml (median lethal dose estimated at earlier), 5 days after treatment individuals are sacrificed. After dissection cuticles are recovered and then subjected to histological sections. The histological technique followed is that of Martoja Martoja-Pierson (1967). Microscopic observation revealed alterations in the architecture of the cuticule which leads to disorganization of cell layers.

Keywords: biopesticide, cuticle, desert locust, toxicity

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1131 Computer-Aided Detection of Simultaneous Abdominal Organ CT Images by Iterative Watershed Transform

Authors: Belgherbi Aicha, Hadjidj Ismahen, Bessaid Abdelhafid

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Interpretation of medical images benefits from anatomical and physiological priors to optimize computer-aided diagnosis applications. Segmentation of liver, spleen and kidneys is regarded as a major primary step in the computer-aided diagnosis of abdominal organ diseases. In this paper, a semi-automated method for medical image data is presented for the abdominal organ segmentation data using mathematical morphology. Our proposed method is based on hierarchical segmentation and watershed algorithm. In our approach, a powerful technique has been designed to suppress over-segmentation based on mosaic image and on the computation of the watershed transform. Our algorithm is currency in two parts. In the first, we seek to improve the quality of the gradient-mosaic image. In this step, we propose a method for improving the gradient-mosaic image by applying the anisotropic diffusion filter followed by the morphological filters. Thereafter, we proceed to the hierarchical segmentation of the liver, spleen and kidney. To validate the segmentation technique proposed, we have tested it on several images. Our segmentation approach is evaluated by comparing our results with the manual segmentation performed by an expert. The experimental results are described in the last part of this work.

Keywords: anisotropic diffusion filter, CT images, morphological filter, mosaic image, simultaneous organ segmentation, the watershed algorithm

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1130 Modeling Optimal Lipophilicity and Drug Performance in Ligand-Receptor Interactions: A Machine Learning Approach to Drug Discovery

Authors: Jay Ananth

Abstract:

The drug discovery process currently requires numerous years of clinical testing as well as money just for a single drug to earn FDA approval. For drugs that even make it this far in the process, there is a very slim chance of receiving FDA approval, resulting in detrimental hurdles to drug accessibility. To minimize these inefficiencies, numerous studies have implemented computational methods, although few computational investigations have focused on a crucial feature of drugs: lipophilicity. Lipophilicity is a physical attribute of a compound that measures its solubility in lipids and is a determinant of drug efficacy. This project leverages Artificial Intelligence to predict the impact of a drug’s lipophilicity on its performance by accounting for factors such as binding affinity and toxicity. The model predicted lipophilicity and binding affinity in the validation set with very high R² scores of 0.921 and 0.788, respectively, while also being applicable to a variety of target receptors. The results expressed a strong positive correlation between lipophilicity and both binding affinity and toxicity. The model helps in both drug development and discovery, providing every pharmaceutical company with recommended lipophilicity levels for drug candidates as well as a rapid assessment of early-stage drugs prior to any testing, eliminating significant amounts of time and resources currently restricting drug accessibility.

Keywords: drug discovery, lipophilicity, ligand-receptor interactions, machine learning, drug development

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1129 Theoretical Insight into Ligand Free Manganese Catalyzed C-O Coupling Protocol for the Synthesis of Biaryl Ethers

Authors: Carolin Anna Joy, Rohith K. R, Rehin Sulay, Parvathy Santhoshkumar, G.Anil Kumar, Vibin Ipe Thomas

Abstract:

Ullmann coupling reactions are gaining great relevance owing to their contribution in the synthesis of biologically and pharmaceutically important compounds. Palladium and many other heavy metals have proven their excellent ability in coupling reaction, but the toxicity matters. The first-row transition metal also possess toxicity, except in the case of iron and manganese. The suitability of manganese as a catalyst is achieving great interest in oxidation, reduction, C-H activation, coupling reaction etc. In this presentation, we discuss the thermo chemistry of ligand free manganese catalyzed C-O coupling reaction between phenol and aryl halide for the synthesis of biaryl ethers using Density functional theory techniques. The mechanism involves an oxidative addition-reductive elimination step. The transition state for both the step had been studied and confirmed using Intrinsic Reaction Coordinate (IRC) calculation. The barrier height for the reaction had also been calculated from the rate determining step. The possibility of other mechanistic way had also been studied. To achieve further insight into the mechanism, substrate having various functional groups is considered in our study to direct their effect on the feasibility of the reaction.

Keywords: Density functional theory, Molecular Modeling, ligand free, biaryl ethers, Ullmann coupling

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1128 Correlation of P53 Gene Expression With Serum Alanine Transaminase Levels and Hepatitis B Viral Load in Cirrhosis and Hepatocellular Carcinoma Patients

Authors: Umme Shahera, Saifullah Munshi, Munira Jahan, Afzalun Nessa, Shahinul Alam, Shahina Tabassum

Abstract:

The development of HCC is a multi-stage process. Several extrinsic factors, such as aflatoxin, HBV, nutrition, alcohol, and trace elements are thought to initiate or/and promote the hepatocarcinogenesis. Alteration of p53 status is an important intrinsic factor in this process as p53 is essential for preventing inappropriate cell proliferation and maintaining genome integrity following genotoxic stress. This study was designed to assess the correlation of p53 gene expression with HBV-DNA and serum Alanine transaminase (ALT) in patients with cirrhosis and HCC. The study was conducted among 60 patients. The study population were divided into four groups (15 in each groups)-HBV positive cirrhosis, HBV negative cirrhosis, HBV positive HCC and HBV negative HCC. Expression of p53 gene was observed using real time PCR. P53 gene expressions in the above mentioned groups were correlated with serum ALT level and HBV viral load. p53 gene was significantly higher in HBV-positive patients with HCC than HBV-positive cirrhosis. Similarly, the expression of p53 was significantly higher in HBV-positive HCC than HBV-negative HCC patients. However, the expression of p53 was reduced in HBV-positive cirrhosis in comparison with HBV-negative cirrhosis. P53 gene expression in liver was not correlated with the serum levels of ALT in any of the study groups. HBV- DNA load also did not correlated with p53 gene expression in HBV positive HCC and HBV positive cirrhosis patients. This study shows that there was no significant change with the expression of p53 gene in any of the study groups with ALT level or viral load, though differential expression of p53 gene were observed in cirrhosis and HCC patients.

Keywords: P53, ALT, HBV-DNA, liver cirrhosis, hepatocellular carcinoma

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1127 Effects of Boldenone Injections and Endurance Exercise on Hepatocyte Morphologic Damages in Male Wistar Rats

Authors: Seyyed Javad Ziaolhagh

Abstract:

Background: The purpose of present study was to investigate, the effects of anabolic steroid Boldenone (BOL) with eight weeks of resistance training on structural changes in rat liver. Method: 21 Male adult Wistar rats, 12 weeks old and 228/53±7/94 g initial body weight were randomly assigned to three groups: group1: Control+ Placebo (C), group2: training+ Placebo (T), group3: Boldenone intramuscular injections 5mg/kg (B). The endurance training protocol consisted three exercise sessions weekly started by a 30-minute run with the speed of 12 m/min and lasted by 60min run with the speed of 30 m/min in 8 weeks. At the end of the experiment, for light microscopic study Slides were prepared. Results: Sections stained of rat's livers showed no any cell degeneration and cytoplasmic lipid vacuoles in all groups, but few samples were seen. Indeed, congested blood sinusoids, cell infiltration and degeneration were seen in the Boldenone-treated group. Hepatotoxic effects were severe in group treatment received 5 mg/kg and directly depended on the doses. Indeed, training group was no any hepatocyte degeneration, inflammation and congestion. Conclusion: The present results showed that BOL has a marked adverse effect on the liver tissue, even with low– dose and endurance training. As a result, athletes should aware of Boldenone dosage consumption.

Keywords: anabolic androgenic steroids, Boldenone, blood congestion, cellular inflammation, cellular degeneration, lipid vocuolations, endurance training

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1126 3D Interactions in Under Water Acoustic Simulationseffect of Green Synthesized Metal Nanoparticles on Gene Expression in an In-Vitro Model of Non-alcoholic Steatohepatitis

Authors: Nendouvhada Livhuwani Portia, Nicole Sibuyi, Kwazikwakhe Gabuza, Adewale Fadaka

Abstract:

Metabolic dysfunction-associated liver disease (MASLD) is a chronic condition characterized by excessive fat accumulation in the liver, distinct from conditions caused by alcohol, viral hepatitis, or medications. MASLD is often linked with metabolic syndrome, including obesity, diabetes, hyperlipidemia, and hypertriglyceridemia. This disease can progress to metabolic dysfunction-associated steatohepatitis (MASH), marked by liver inflammation and scarring, potentially leading to cirrhosis. However, only 43-44% of patients with steatosis develop MASH, and 7-30% of those with MASH progress to cirrhosis. The exact mechanisms underlying MASLD and its progression remain unclear, and there are currently no specific therapeutic strategies for MASLD/MASH. While anti-obesity and anti-diabetic medications can reduce progression, they do not fully treat or reverse the disease. As an alternative, green-synthesized metal nanoparticles (MNPs) are emerging as potential treatments for liver diseases due to their anti-diabetic, anti-inflammatory, and anti-obesity properties with minimal side effects. MNPs like gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) have been shown to improve metabolic processes by lowering blood glucose, body fat, and inflammation. The study aimed to explore the effects of green-synthesized MNPs on gene expression in an in vitro model of MASH using C3A/HepG2 liver cells. The MASH model was created by exposing these cells to free fatty acids (FFAs) followed by lipopolysaccharide (LPS) to induce inflammation. Cell viability was assessed with the Water-Soluble Tetrazolium (WST)-1 assay, and lipid accumulation was measured using the Oil Red O (ORO) assay. Additionally, mitochondrial membrane potential was assessed by the tetramethyl rhodamine, methyl ester (TMRE) assay, and inflammation was measured with an Enzyme-Linked Immunosorbent Assay (ELISA). The study synthesized AuNPs from Carpobrotus edulis fruit (CeF) and avocado seed (AvoSE) and AgNPs from Salvia africana-lutea (SAL) using optimized conditions. The MNPs were characterized by UV-Vis spectrophotometry and Dynamic Light Scattering (DLS). The nanoparticles were tested at various concentrations for their impact on the C3A/HepG2-induced MASH model. Among the MNPs tested, AvoSE-AuNPs showed the most promise. They reduced cell proliferation and intracellular lipid content more effectively than CeFE-AuNPs and SAL-AgNPs. Molecular analysis using real-time polymerase chain reaction revealed that AvoSE-AuNPs could potentially reverse MASH effects by reducing the expression of key pro-inflammatory and metabolic genes, including tumor necrosis factor-alpha (TNF-α), Fas cell surface death receptor (FAS), Peroxisome proliferator-activated receptor (PPAR)-α, PPAR-γ, and Sterol regulatory element-binding protein (SREBPF)-1. Further research is needed to confirm the molecular mechanisms behind the effects of these MNPs and to identify the specific phytochemicals responsible for their synthesis and bioactivities.

Keywords: gold nanoparticles, green nanotechnology, metal nanoparticles, obesity

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1125 Inhibitory Impacts of Fulvic Acid-Coated Iron Oxide Nano Particles on the Amyloid Fibril Aggregations

Authors: Dalia Jomehpour, Sara Sheikhlary, Esmaeil Heydari, Mohammad Hossien Majles Ara

Abstract:

In this study, we report fulvic acid-coated iron oxide nanoparticles of 10.7 ± 2.7 nm size, which serve to inhibit amyloid fibrillation formation. Although the effect of fulvic acid on tau fibrils was investigated, to our best knowledge, its inhibitory impacts on amyloid aggregation formation have been assessed neither in-vitro nor in-vivo. On the other hand, iron oxide nanoparticles exhibit anti-amyloid activity on their own. This study investigates the inhibitory effect of fulvic acid coated iron oxide nanoparticles on amyloid aggregations formed from the commonly used in-vitro model, lysozyme from chicken egg white. FESEM, XRD, and FTIR characterization confirmed that fulvic acid was coated onto the surface of the nanoparticles. The inhibitory effects of the fulvic acid coated iron oxide nanoparticles were verified by Thioflavin T assay, circular dichroism (CD), and FESEM analysis. Furthermore, the toxicity of the nanoparticles on the neuroblastoma SH-SY5Y human cell line was assessed through an MTT assay. Our results indicate that fulvic acid coated iron oxide nanoparticles can efficiently inhibit the formation of amyloid aggregations while exhibiting negligible in-vitro toxicity; thus, they can be used as anti-amyloid agents in the development of the potential drug for neurodegenerative diseases.

Keywords: Alzheimer’s disease, fulvic acid coated iron oxide nanoparticles, fulvic acid, amyloid inhibitor, polyphenols

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1124 Analysis of the Presence of Alkylglycerols by Gas Chromatography in Ostrich Oil

Authors: Luana N. Cardozo, Debora A. S. Coutinho, Fabiola Lagher, Bruno J. G. Silva, Ivonilce Venture, Mainara Tesser, Graciela Venera

Abstract:

Ostrich oil is used as food in Brazil, and it has been the subject of scientific research because it contains essential fatty acids (Omega 3, 6, 7, and 9), which provide benefits to human health. Alkylglycerols are lipid ethers consisted of a saturated or unsaturated hydrocarbon chain joined by ether-type bonding to one of the glycerol hydroxyls. It is known that supplementation with alkylglycerols can act significantly on the functioning of immune system cells, both in pathological situations and in homeostasis. Objective: Analyze the presence of alkylglycerols in ostrich oil. Methods: The ostrich oil was bought from an industry that manufactures the product for sale as food, located in Mirante da Serra, northern Brazil. The samples were sent for analysis to the chemistry department of the Federal University of Paraná, where they were analyzed by the gas chromatography method. Results: The analysis of the ostrich oil presented alkylglycerols in area 514505154. Comparison, it is possible to observe that shark liver oil contains the area 26190196, and the difference between both is highly significant. Conclusion: The importance of alkylglycerol supplementation for the immune system is known. The analysis of the results made it possible to verify the presence of alkylglycerols in the ostrich oil, which is five times higher than in the shark liver oil, that would be the largest source food, but was surpassed by the ostrich oil until the present time. The present study emphasizes that ostrich oil can be considered a food source of alkylglycerols and may play a promising role in the immune system because it contains such substance, but further studies are needed to prove its performance in the body.

Keywords: ostrich oil, nutritional composition, alkylglycerols, food

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1123 Bioactivities and Phytochemical Studies of Acrocarpus fraxinifolius Bark Wight and Arn

Authors: H. M. El-Rafie, A. H. Abou Zeid, R. S. Mohammed, A. A. Sleem

Abstract:

Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to investigate the phytoconstituents of the petroleum ether extract (PEE) of Acrocarpus fraxinofolius bark by Gas chromatography coupled with mass spectrometry (GC/MS) analysis of its fractions (fatty acid and unsaponifiable matter). Concerning this, identification of 52 compounds constituting 97.03 % of the total composition of the unsaponifiable matter fraction. Cycloeucalenol was found to be the major compound representing 32.52% followed by 4a, 14a-dimethyl-A8~24(28)-ergostadien (26.50%) and ß-sitosterol(13.74%), furthermore Gas liquid chromatography (GLC) analysis of the sterol fraction revealed the identification of cholesterol (7.22 %), campesterol (13.30 %), stigmasterol (10.00 %) and β - sitosterol (69.48 %). Meanwhile, the identification of 33 fatty acids representing 90.71% of the total fatty acid constituents. Methyl-9,12-octadecadienoate (40.39%) followed by methyl hexadecanoate (23.64%) were found to be the major compounds. On the other hand, column chromatography and Thin layer chromatography (TLC) fractionation of PEE separate the triterpenoid: 21β-hydroxylup-20(29)-en-3-one and β- amyrin which were structurally identified by spectroscopic analysis (NMR, MS and IR). PEE has been biologically evaluated for 1: management of diabetes in alloxan induced diabetic rats 2: cytotoxic activity against four human tumor cell lines (Cervix carcinoma cell line[HELA], Breast carcinoma cell line [MCF7], Liver carcinoma cell line[HEPG2] and Colon carcinoma cell line[HCT-116] 3: hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin (100% potency). It also showed a significant anti-proliferative activity against MCF-7 (IC50= 2.35µg), Hela(IC50=3.85µg) and HEPG-2 (IC50= 9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity was evidenced by significant decrease in liver function enzymes, i.e. AST, ALT and ALP by (29.18%, 28.26%, and 34.11%, respectively using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. This study was performed for the first time on the bark of this species.

Keywords: Acrocarpus fraxinofolius, antidiabetic, cytotoxic, hepatoprotective

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1122 The Investigation of Effectiveness of Different Concentrations of the Mycotoxin Detoxification Agent Added to Broiler Feed, in the Presence of T-2 Toxin, on Performance, Organ Mass and the Residues T-2 Toxin and His Metabolites in the Broiler Tissues

Authors: Jelena Nedeljković Trailović, Marko Vasiljević, Jog Raj, Hunor Farkaš, Branko Petrujkić, Stamen Radulović, Gorana Popvić

Abstract:

The experiment was performed on a total of 99 one-day-old broilers of Cob 500 provenance, which were divided into IX equal groups. Broilers of the E-I group were fed 0.25 mg T-2 toxin/kg feed, E-II and E-III groups 0.25 mg T-2 toxin/kg feed with the addition of 1 kg/t and 3 kg/t of the mycotoxin detoxification agent MDA, respectively. The E-IV group received 1 mg of T-2 toxin/kg of feed, and the broilers of E-V and E-VI groups received 1 mg of T-2 toxin/kg of feed with the addition of 1 kg/t and 3 kg/t of the MDA detoxification preparation, respectively. The E-VII group received commercial feed without toxins and additives, the E-VIII and E-IX groups received feed with 1kg/t and 3kg/t of the MDA detoxification preparation. The trial lasted 42 days. Observing the results obtained on the 42nd day of the experiment, we can conclude that the change in the absolute mass of the spleen occurred in the broilers of the E-IV group (1.66±0.14)g, which was statistically significantly lower compared to the broilers of the E-V and E-VI groups (2.58±0.15 and 2.68±0.23)g. Heart mass was significantly statistically lower in broilers of group E-IV (9.1±0.38)g compared to broilers of group E-V and E-VI (12.23±0.5 and 11.43±0.51)g. It can be concluded that the broilers that received 1 kg/t and 3 kg/t of the detoxification preparation had an absolute mass of organs within physiological limits. Broilers of the E-IV group achieved the lowest BM during the experiment (on the 42nd day of the experiment 1879±52.73)g, they were significantly statistically lower than the BW of broilers of all experimental groups. This trend is observed from the beginning to the end of the experiment. The protective effect of the detoxification preparation can be seen in broilers of the E-V group, that had a significantly statistically higher BM on the 42nd day of the experiment (2225±58.81)g compared to broilers of group E-IV. Broilers of E-VIII group (2452±46.71) g, which received commercial feed with the addition of 1 kg/t MDA preparation, had the highest BMI at the end of the experiment. At the end of the trial on the 42nd day, blood samples were collected from broilers of the experimental groups that received T-2 toxin and MR detoxification preparations in different concentrations. Also, liver and breast musculature samples were collected for testing for the presence and content of T-2 toxin, HT-2 toxin, T-2 tetraol and T-2 triol. Due to very rapid elimination from the blood, no remains of T-2 toxin and its metabolites were detected in the blood of broilers of groups E-I to E-VI. In the breast muscles, T-2 toxin residues below LoQ < 0.2 (μg/kg) were detected in all groups that received T-2 toxin in food, the highest value was recorded in the E-IV group (0.122 μg/kg and the lowest in E -VI group 0.096 μg/kg). No T-2 toxin residues were detected in the liver. Remains of HT-2 were detected in the breast muscles and livers of broilers from E-IV, E-V and E-VI groups, LoQ < 1 (μg/kg); for the breast muscles: 0.054, 0.044 and 0.041 μg/kg, and for the liver: 0.473, 0.231 and 0.185 μg/kg. Summing up all the results, a partial protective effect of the detoxification preparation, added to food in the amount of 1kg/t, can be seen.

Keywords: T-2 toxin, bloiler, MDA, mycotoxuns

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1121 Therapeutic Drug Monitoring by Dried Blood Spot and LC-MS/MS: Novel Application to Carbamazepine and Its Metabolite in Paediatric Population

Authors: Giancarlo La Marca, Engy Shokry, Fabio Villanelli

Abstract:

Epilepsy is one of the most common neurological disorders, with an estimated prevalence of 50 million people worldwide. Twenty five percent of the epilepsy population is represented in children under the age of 15 years. For antiepileptic drugs (AED), there is a poor correlation between plasma concentration and dose especially in children. This was attributed to greater pharmacokinetic variability than adults. Hence, therapeutic drug monitoring (TDM) is recommended in controlling toxicity while drug exposure is maintained. Carbamazepine (CBZ) is a first-line AED and the drug of first choice in trigeminal neuralgia. CBZ is metabolised in the liver into carbamazepine-10,11-epoxide (CBZE), its major metabolite which is equipotent. This develops the need for an assay able to monitor the levels of both CBZ and CBZE. The aim of the present study was to develop and validate a LC-MS/MS method for simultaneous quantification of CBZ and CBZE in dried blood spots (DBS). DBS technique overcomes many logistical problems, ethical issues and technical challenges faced by classical plasma sampling. LC-MS/MS has been regarded as superior technique over immunoassays and HPLC/UV methods owing to its better specificity and sensitivity, lack of interference or matrix effects. Our method combines advantages of DBS technique and LC-MS/MS in clinical practice. The extraction process was done using methanol-water-formic acid (80:20:0.1, v/v/v). The chromatographic elution was achieved by using a linear gradient with a mobile phase consisting of acetonitrile-water-0.1% formic acid at a flow rate of 0.50 mL/min. The method was linear over the range 1-40 mg/L and 0.25-20 mg/L for CBZ and CBZE respectively. The limit of quantification was 1.00 mg/L and 0.25 mg/L for CBZ and CBZE, respectively. Intra-day and inter-day assay precisions were found to be less than 6.5% and 11.8%. An evaluation of DBS technique was performed, including effect of extraction solvent, spot homogeneity and stability in DBS. Results from a comparison with the plasma assay are also presented. The novelty of the present work lies in being the first to quantify CBZ and its metabolite from only one 3.2 mm DBS disc finger-prick sample (3.3-3.4 µl blood) by LC-MS/MS in a 10 min. chromatographic run.

Keywords: carbamazepine, carbamazepine-10, 11-epoxide, dried blood spots, LC-MS/MS, therapeutic drug monitoring

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1120 Association of Transmission Risk Factors Among HCV-infected Bangladeshi Patients With Different Genotypes

Authors: Nahida Sultana

Abstract:

Globally, an estimated 58 million people have chronic hepatitis C virus infection, with about 1.5 million new infections occurring per year. The hepatitis C virus is a blood-borne virus, and most infections occur through exposure to blood from unsafe injection practices, unsafe health care, unscreened blood transfusion, injection drug use, and sexual practices that lead to exposure to blood. Hepatitis C virus (HCV) causes chronic infections that mainly affect the liver leading to liver diseases. This study aimed to determine whether there is any significant association between HCV transmission risk factors in relation to genotypes in HCV-infected Bangladeshi patients. After quantification of HCV viral load, 36 samples were randomly selected for HCV genotyping and risk factor measurement. A greater proportion of genotype 1 (p > 0.05) patients (40%) underwent blood transfusion compared to patients (22.6%) with genotype 3 infections. More genotype 1 patient underwent surgery and invasive procedures (20%), and rather than those with genotype 3 patients (16.1%). The history of IDUs (25.8%) and sexual exposure (3.2%) are only prevalent in genotype 3 patients and absent in patients with genotype 1 (p >0.05). There was no significant statistical difference found in HCV transmission risk factors (blood transfusion, IDUs, Surgery& interventions, sexual transmission) between patients infected with genotypes 1 and 3. In HCV infection, genotype may have no relation to transmission risk factors among Bangladeshi patients.

Keywords: HCV genotype, alanine aminotransferase (ALT), HCV viral load, IDUs

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1119 Comparison of Phenolic and Urushiol Contents of Different Parts of Rhus verniciflua and Their Antimicrobial Activity

Authors: Jae Young Jang, Jong Hoon Ahn, Jae-Woong Lim, So Young Kang, Mi Kyeong Lee

Abstract:

Rhus verniciflua is commonly known as a lacquer tree in Korea. Stem barks of R. verniciflua have been used as an immunostimulator in traditional medicine. It contains phenolic compounds and is known for diverse biological activities such as antioxidant and antimicrobial activity. However, it also causes allergic dermatitis due to urushiols derivatives. For the development of active natural resources with less toxicity, the content of phenolic compounds and urushiols of different parts of R. verniciflua such as stem barks, lignum and leaves were quantitated by colorimetric assay and HPLC analysis. The urushiols content were the highest in stem barks, and followed by leaves. The lignum contained trace amount of urushiols. The phenolic contents, however, were the most abundant in lignum, and followed by leaves and stem barks. These results clear showed that the content of urushiols and phenolic differs depending on the parts of R. verniciflua. Antimicrobial activity of different parts of R. verniciflua against fish pathogenic bacteria was also investigated using Edwardsiella tarda. Lignum of R. verniciflua was the most effective in antimicrobial activity against E. tarda and phenolic constituents are suggested to be active constituents for activity. Taken together, phenolic compounds are responsible for antimicrobial activity of R. verniciflua. The lignum of R. verniciflua contains high content of phenolic compounds with less urushiols, which suggests efficient antimicrobial activity with less toxicity. Therefore, lignum of R. verniciflua are suggested as good sources for antimicrobial activity against fish bacterial diseases.

Keywords: different parts, phenolic compounds, Rhus verniciflua, urushiols

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1118 Scanning Electron Microscopy of the Erythrocytes of Channa punctatus (Bloch) Exposed to Mercuric Chloride

Authors: Shweta Maheshwari, Anish Dua

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Hematological changes reflect the adverse effects of heavy metals on fish. Hematology is a valuable tool to evaluate pathological condition of the fish. It helps in diagnosing the structural and functional status of fish exposed to toxicants. Morphological alteration in erythrocytes due to environmental stress can be studied through ultra-structural analysis. The aim of the present study was to assess the toxicity of mercuric chloride on red blood cells of an air breathing fish, Channa punctatus. Fish were subjected to chronic experiments using three sublethal concentration of mercuric chloride (0.020mg/L, 0.027mg/L, 0.040mg/L) for a period of 15, 30 and 60 days. Exposed fish of all the three concentrations were subjected to a recovery period of 30 days. A control was maintained in tap water simultaneously. For SEM analysis, blood from caudal vein of fish was taken and examined at an accelerating voltage of 20kV. Scanning electron micrographs revealed elliptical shaped erythrocytes of control fish. Alterations in the erythrocyte morphology such as presence of spherocytes, membrane internalization, crenation of membrane and development of lobopodial projections were observed in the exposed fish. The study revealed that ultra-structural analysis appears to be a sensitive method to evaluate the toxicity of various toxicants to fish.

Keywords: Channa punctatus, erythrocytes, mercuric chloride, scanning electron microscopy

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1117 Synthesis and Biological Evaluation of Pyridine Derivatives as Antimicrobial Agents

Authors: Dagim Ali Hussen, Adnan A. Bekhit, Ariaya Hymete

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In this study, several pyridine derivatives were synthesized and evaluated for their in vitro antimicrobial activity against gram-positive bacteria (S. aureus and B. Cereus), gram-negative bacteria (P. aeruginosa and E. coli) and fungus (C. albican and A niger). The intermediate chalcone derivative 2a,b was synthesized by condensation of pyrazole aldehydes 1a,b with acetophenone in alcoholic KOH. Cyclization of 2a,b with ethyl cyanoacetate ad ammonium acetate resulted in pyridine carbonitrile derivatives 3a,b. Furthermore, condensation of pyridine-4-carboxaldeyhe with different amino-derivatives gave rise to pyridine derivatives 5a,b, 6a,b. The oxadiazole derivative 7a was prepared by cyclization of 6a with acetic anhydride. Characterization of the synthesized compound was performed using IR, 1H NMR, 13C NMR spectra and elemental microanalyses. The antimicrobial results revealed that compounds 5a, 6b and 7a exhibited half fold antibacterial activity compared to ampicillin, against B. cereus. On the other hand, compound 3b showed an equivalent activity compared to miconazole against candida albican (CANDAL 03) and to clotrimazole against the clinical isolate candida albican 6647. Moreover, this compound 3b was further tested for its acute toxicity profile. The results showed that oral LD50 is more that 300 mg/kg and parentral LD50 is more than 100 mg/kg. Compound 3b is a good candidate for antifungal agent with good toxicity profile, and deserves more chemical derivatization and clinical study.

Keywords: antifungal, antimicrobial, Candida albican, pyridine

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