Search results for: zoospore inhibition assay

Authors: Nadine Wiesmann, Melanie Viel, Christoph Buhr, Rachel Tanner, Wolfgang Tremel, Juergen Brieger

Abstract:

Recidivation of tumors and the development of resistances against the classical anti-tumor approaches represent a major challenge we face when treating cancer. In order to master this challenge, we are in desperate need of new treatment options beyond the beaten tracks. Zinc oxide nanoparticles (ZnO NPs) represent such an innovative approach. Zinc oxide is characterized by a high level of biocompatibility, concurrently ZnO NPs are able to exert anti-tumor effects. By concentration of the nanoparticles at the tumor site, tumor cells can specifically be exposed to the nanoparticles while low zinc concentrations at off-target sites are tolerated well and can be excreted easily. We evaluated the toxicity of ZnO NPs in vitro with the help of immortalized tumor cell lines and primary cells stemming from healthy tissue. Additionally, the Chorioallantoic Membrane Assay (CAM Assay) was employed to gain insights into the in vivo behavior of the nanoparticles. We could show that ZnO NPs interact with tumor cells as nanoparticulate matter. Furthermore, the extensive release of zinc ions from the nanoparticles nearby and within the tumor cells results in overload with zinc. Beyond that, ZnO NPs were found to further the generation of reactive oxygen species (ROS). We were able to show that tumor cells were more prone to the toxic effects of ZnO NPs at intermediate concentrations compared to fibroblasts. With the help of ZnO NPs covered by a silica shell in which FITC dye was incorporated, we were able to track ZnO NPs within tumor cells as well as within a whole organism in the CAM assay after injection into the bloodstream. Depending on the applied concentrations, selective tumor cell killing seems feasible. Furthermore, the combinational treatment of tumor cells with radiotherapy and ZnO NPs shows promising results. Still, further investigations are needed to gain a better understanding of the interaction between ZnO NPs and the human body to be able to pave the way for their application as an innovative anti-tumor agent in the clinics.

Keywords: metal oxide nanoparticles, nanomedicine, overcome resistances against classical treatment options, zinc oxide nanoparticles

Procedia PDF Downloads 119

Authors: Harald Lindorfer, Bettina Frauz, Dietmar Ramhold

Abstract:

Next to the well-known inhibitors in anaerobic digestion like ammonia, antibiotics or disinfectants, the number of process failures connected with mould growth in the feedstock increased significantly in the last years. It was assumed that mycotoxins are the cause of the negative effects. The financial damage to plants associated with these process failures is considerable. The aim of this study was to find a way of predicting the failures and furthermore strategies for a fast process recovery. In a first step, mould-contaminated feedstocks causing process failures in full-scale digesters were sampled and analysed on mycotoxin content. A selection of these samples was applied to biological inhibition tests. In this test, crystalline cellulose is applied in addition to the feedstock sample as standard substrate. Affected digesters were also sampled and analytical process data as well as operational data of the plants were recorded. Additionally, different mycotoxin substances, Deoxynivalenol, Zearalenon, Aflatoxin B1, Mycophenolic acid and Citrinin, were applied as pure substances to lab-scale digesters, individually and in various combinations, and effects were monitored. As expected, various mycotoxins were detected in all of the mould-contaminated samples. Nevertheless, inhibition effects were observed with only one of the collected samples, after applying it to an inhibition test. With this sample, the biogas yield of the standard substrate was reduced by approx. 20%. This result corresponds with observations made on full-scale plants. However, none of the tested mycotoxins applied as pure substance caused a negative effect on biogas production in lab scale digesters, neither after application as individual substance nor in combination. The recording of the process data in full-scale plants affected by process failures in most cases showed a severe accumulation of fatty acids alongside a decrease in biogas production and methane concentration. In the analytical data of the digester samples, a typical distribution of fatty acids with exceptionally high acetic acid concentrations could be identified. This typical fatty acid pattern can be used as a rapid identification parameter pointing to the cause of the process troubles and enable a fast implication of countermeasures. The results of the study show that more attention needs to be paid to feedstock storage and feedstock conservation before their application to anaerobic digesters. This is all the more important since first studies indicate that the occurrence of mycotoxins will likely increase in Europe due to the ongoing climate change.

Keywords: Anaerobic digestion, Biogas, Feedstock conservation, Fungal mycotoxins, Inhibition, process failure

Procedia PDF Downloads 113

Authors: Reena Murali, David Peter S.

Abstract:

The small interfering RNA (siRNA) alters the regulatory role of mRNA during gene expression by translational inhibition. Recent studies shows that up regulation of mRNA cause serious diseases like Cancer. So designing effective siRNA with good knockdown effects play an important role in gene silencing. Various siRNA design tools had been developed earlier. In this work, we are trying to analyze the existing good scoring second generation siRNA predicting tools and to optimize the efficiency of siRNA prediction by designing a computational model using Artificial Neural Network and whole stacking energy (ΔG), which may help in gene silencing and drug design in cancer therapy. Our model is trained and tested against a large data set of siRNA sequences. Validation of our results is done by finding correlation coefficient of experimental versus observed inhibition efficacy of siRNA. We achieved a correlation coefficient of 0.727 in our previous computational model and we could improve the correlation coefficient up to 0.753 when the threshold of whole tacking energy is greater than or equal to -32.5 kcal/mol.

Keywords: artificial neural network, double stranded RNA, RNA interference, short interfering RNA

Procedia PDF Downloads 515

Authors: Hoo Cheol Lee, Dae Seung Kim, Sang Myung Jung, Gwang Heum Yoon, Hwa Sung Shin

Abstract:

Loss of bone tissue can occur due to a bone tissue disease and aging or fracture. Renewable formation of bone is mainly made by its differentiation and metabolism. For this reason, osteoblasts have been studied for regeneration of bone tissue. So, tissue engineering has attracted attention as a recovery means. In tissue engineering, a particularly important factor is a scaffold that supports cell growth. For osteoblast scaffold, we used the cellulose nanowhisker (CNW) extracted from marine organism. CNW is one of an abundant material obtained from a number of plants and animals. CNW is polymer consisting of monomer cellulose and this composition offers biodegradability and biocompatibility to CNW. Mechanical strength of CNW is superior to the existing natural polymers. In addition, substances of marine origin have a low risk of secondary infection by bacteria and pathogen in contrast with those of land-derived. For evaluating its suitability as an osteoblast scaffold, we fabricate CNW film for osteoblast culture and performed the MTT assay and ALP assay to confirm its cytotoxicity and effect on differentiation. Taking together these results, we assessed CNW is a potential candidate of a material for bone tissue regeneration.

Keywords: bone regeneration, cellulose nanowhisker, marine derived material, osteoblast

Procedia PDF Downloads 335

Authors: Zhaoguo Liu, Cunsi Shen, Yin Lu

Abstract:

Growing evidence has shown that menthol has potent anticancer activity in various human cancers. However, its effect on skin cancer remains largely unknown. In the present study, we investigated the chemopreventive potential of menthol against 7, 12-dimethylbenz[a] anthracene(DMBA)/12-O-tetradecanoylphorbol 13-acetate (TPA)-induced skin tumorigenesis in ICR mice. Our results showed that menthol significantly inhibited TPA-induced inflammatory responses and pro-inflammatory cytokine release. We also found that menthol treatment significantly inhibited TPA-induced lipid peroxidation (LPO), mouse UDP-glucumno-syltransferase (UGT), mouse NADH Dehydrogenase, Quinone 1 (NQO1) release. Furthermore, we found menthol treatment significantly inhibited the tumor incidence and number of tumors (P < 0.001). Interestingly, we observed that menthol treatment significantly inhibited TPA-induced altered activity of NF-κB in skin tumor. Consistently, menthol-treated tumors also showed significantly suppressed the Ras-Raf-ERK signaling pathway. Thus, our results suggest that menthol inhibits DMBA/TPA-induced skin tumorigenesis by attenuating the Ras and inhibiting NF-κB activity via inhibition of inflammation responses and pro-inflammatory cytokine release.

Keywords: DMBA/TPA, NF-κB, Ras-Raf-ERK, skin tumorigenesis

Procedia PDF Downloads 302

Authors: Ogemdi Chinwendu Anika, Anna Strzelecka, Yadira Bajón-Fernández, Raffaella Villa

Abstract:

Anaerobic digestion (AD) has been used since time in memorial to take care of organic wastes in the environment, especially for sewage and wastewater treatments. Recently, the rising demand/need to increase renewable energy from organic matter has caused the AD substrates spectrum to expand and include a wider variety of organic materials such as agricultural residues and farm manure which is annually generated at around 140 billion metric tons globally. The problem, however, is that agricultural wastes are composed of materials that are heterogeneous and too difficult to degrade -particularly lignin, that make up about 0–40% of the total lignocellulose content. This study aimed to evaluate the impact of varying concentrations of lignin on biogas yields and their subsequent response to a commercial yeast-based bioadditive in batch anaerobic digesters. The experiments were carried out in batches for a retention time of 56 days with different lignin concentrations (200 mg, 300 mg, 400 mg, 500 mg, and 600 mg) treated to different conditions to first determine the concentration of the bioadditive that was most optimal for overall process improvement and yields increase. The batch experiments were set up using 130 mL bottles with a working volume of 60mL, maintained at 38°C in an incubator shaker (150rpm). Digestate obtained from a local plant operating at mesophilic conditions was used as the starting inoculum, and commercial kraft lignin was used as feedstock. Biogas measurements were carried out using the displacement method and were corrected to standard temperature and pressure using standard gas equations. Furthermore, the modified Gompertz equation model was used to non-linearly regress the resulting data to estimate gas production potential, production rates, and the duration of lag phases as indicatives of degrees of lignin inhibition. The results showed that lignin had a strong inhibitory effect on the AD process, and the higher the lignin concentration, the more the inhibition. Also, the modelling showed that the rates of gas production were influenced by the concentrations of the lignin substrate added to the system – the higher the lignin concentrations in mg (0, 200, 300, 400, 500, and 600) the lower the respective rate of gas production in ml/gVS.day (3.3, 2.2, 2.3, 1.6, 1.3, and 1.1), although the 300 mg increased by 0.1 ml/gVS.day over that of the 200 mg. The impact of the yeast-based bioaddition on the rate of production was most significant in the 400 mg and 500 mg as the rate was improved by 0.1 ml/gVS.day and 0.2 ml/gVS.day respectively. This indicates that agricultural residues with higher lignin content may be more responsive to inhibition alleviation by yeast-based bioadditive; therefore, further study on its application to the AD of agricultural residues of high lignin content will be the next step in this research.

Keywords: anaerobic digestion, renewable energy, lignin valorisation, biogas

Procedia PDF Downloads 75

Authors: Mthokozisi Simelane

Abstract:

Background: Malaria remains a life-threatening disease in tropical regions despite the advances in the treatment of this disease, it still remains a significant burden as some parasites have become resistant to the currently available drugs. This has created a necessity for the development of alternative, more efficient antimalarial drugs. Warburgia salutaris is a traditional medicinal plant used in malaria treatment by Zulu traditional healers. Materials and methods: The W. salutaris stem-bark was extracted with dichloromethane and the compound was isolated through column chromatography. The compound was identified and characterized by spectroscopic analysis (1H NMR, 13C NMR, IR and MS) and the structure was also confirmed by x-ray crystallography. The anti-plasmodial activity (in vitro) was studied on NF54 Plasmodium falciparum strain (CQS). Cytotoxicity was measured using the MTT assay on HEK239 and HEPG2 cell lines. Docking of Mukaadial acetate was conducted in AutoDock Vina. Structural modifications were conducted in UCSF Chimera and molecular interactions examined in LigPlot. Results: The compound, Mukaadial Acetate showed appreciable inhibition (IC50 0.44±0.10 µg/ml) of the parasite growth and cytotoxicity activity of 0.124±0.109 and 0.199±0.083 (µg/ml) on HEK293 and HEPG2 cells respectively. Molecular docking revealed that Mukaadial Acetate binds to the purine, pyrophosphate and ribose binding sites of the PfHGXPRT with an optimum binding conformation and forms hydrogen bond, steric and hydrophobic interactions with the residues inhabiting the respective binding sites. Conclusion: It is apparent that W. salutaris contains components (including Mukaadial Acetate) that exhibit antimalarial activity. This study scientifically validates the use of this plant in folk medicine.

Keywords: plasmodium falciparum, molecular docking, antimalarial activity, PfHGXPRT, Warburgia salutaris, mukaadial acetate

Procedia PDF Downloads 188

Authors: Roberto Cassino, Valeria Dissette, Carlo Alberto Bignozzi, Daniele Pazzi

Abstract:

Background and Aims: The aim of this work was to investigate, both ‘in vitro’ and ‘in vivo’, if the new SCX technology (SiO2-Ag+Chlorex) can easily defeat infections and it is really more effective than SSD (SilverSulfaDiazine). ‘In vitro’ methods: we tested ‘in vitro’ the effectiveness of both silver materials using a pool of 5 strains: Pseudomonas Aeruginosa, Staphylococcus aureus, Escherichia Coli, Enterococcus hirae and Candida Albicans. 100 µl of this pool have been seeded on Petri dishes and kept for 24 hours in incubation at 37 C°. ‘In vivo’ methods: we enrolled patients with multiple infectious chronic wounds (according with cutting & harding criteria for infection); after a qualitative evaluation of the wounds bacterial population, taking a sample by plug, we included in the study 6 patients for a total of 10 wounds, infected by one or more of the microorganisms used for the ‘in vitro’ test. The protocol consisted of a treatment with a spray powder of SSD every 48 hours for 14 days; in case of worsening we should have to start a new treatment with a spray powder containing silicon dioxide, ionic silver and chlorexidine (SiO2-Ag+Chlorex) every 48 hours for 14 days. We evaluated the number of clinical signs of infection and the disappearance or not of the wound edge erithema. ‘In vitro’ results: SSD demonstrated a wide zone of inhibition within 24 hours, but after 5 days there was no more signs of inhibition; on the contrary SCX had a good inhibition ring that lasted more than 5 days. ‘In vivo’ results: all wounds treated with SSD got worse; the signs of infection increased and the wound edge erithema did not disappear. According with the protocol, we treated then all wounds with SCX and they all improved within the period of observation with complete disappearance of clinical signs of infection and no more wound edge erithema. Conclusions: the study demonstrated the effectiveness of SiO2-Ag+Chlorex, especially in terms of long lasting antimicrobial action. We had the same results ‘in vitro’, so that there has been a perfect correspondence between the laboratory outcomes and the clinical ones.

Keywords: chronic wounds, infections, ionic silver, SSD

Procedia PDF Downloads 315

Authors: Archana. S, Usha Rani. G, Anand Balakrishnan, Sanjana.R, Solomon F, Vijayalakshmi. J

Abstract:

Background: There is evidence that male factors contribute to the infertility of up to 50% of couples, who are evaluated and treated for infertility using advanced assisted reproductive technologies. Genetic abnormalities, including sperm chromosome aneuploidy as well as structural aberrations, are one of the major causes of male infertility. Recent advances in technology expedite the evaluation of sperm aneuploidy. The purpose of the study was to de-termine the prevalence of sperm aneuploidy in infertile males and the degree of association between DNA fragmentation and sperm aneuploidy. Methods: In this study, 75 infertile men were included, and they were divided into four abnormal groups (Oligospermia, Terato-spermia, Asthenospermia and Oligoasthenoteratospermia (OAT)). Men with children who were normozoospermia served as the control group. The Fluorescence in situ hybridization (FISH) method was used to test for sperm aneuploidy, and the Sperm Chromatin Dispersion Assay (SCDA) was used to measure the fragmentation of sperm DNA. Spearman's correla-tion coefficient was used to evaluate the relationship between sperm aneuploidy and sperm DNA fragmentation along with age. P < 0.05 was regarded as significant. Results: 75 partic-ipants' ages varied from 28 to 48 years old (35.5±5.1). The percentage of spermatozoa bear-ing X and Y was determined to be statistically significant (p-value < 0.05) and was found to be 48.92% and 51.18% of CEP X X 1 – nucish (CEP XX 1) [100] and CEP Y X 1 – nucish (CEP Y X 1) [100]. When compared to the rate of DNA fragmentation, it was discovered that infertile males had a greater frequency of sperm aneuploidy. Asthenospermia and OAT groups in sex chromosomal aneuploidy were significantly correlated (p<0.05). Conclusion: Sperm FISH and SCDA assay results showed increased sperm aneuploidy frequency, and DNA fragmentation index in infertile men compared with fertile men. There is a significant relationship observed between sperm aneuploidy and DNA fragmentation in OAT patients. When evaluating male variables and idiopathic infertility, the sperm FISH screening method can be used as a valuable diagnostic tool.

Keywords: ale infertility, dfi (dna fragmentation assay) (scd-sperm chromatin dispersion).art (artificial reproductive technology), trisomy, aneuploidy, fish (fluorescence in-situ hybridization), oat (oligoasthoteratospermia)

Procedia PDF Downloads 42

Authors: Tooba N. Shamsi, Romana Perveen, Sadaf Fatima

Abstract:

Protease Inhibitors (PIs) are widespread in nature, produced by animals, plants and microorganisms. They play vital role in various biological activities by keeping a check on activity of proteases. Present study aims to investigate antioxidant and anti-inflammatory properties of PPI from Cajanus cajan (CCTI) and Phaseolus limensis (LBTI). PPI was purified from C. cajan (PUSA-992) by ammonium sulfate precipitation followed by ion exchange chromatography. The anti-oxidant activity was analyzed by two most common radical scavenging assays of FRAP (ferric reducing antioxidant power) and DPPH (1,1- diphenyl-2-picrylhydrazyl). Also, in-vitro anti-inflammatory activity was evaluated using albumin denaturation assay and membrane stabilization assay at different concentrations. Ascorbic acid and aspirin were used as a standards for antioxidant and anti-inflammatory assays respectively. The PPIs were also checked for antimicrobial activity against a number of bacterial strains. The CCTI and LBTI showed DPPH radical scavenging activity in a concentration–dependent manner with IC50 values 544 µg/ml and 506 µg/ml respectively comparative to ascorbic acid which was 258 µg/ml. Following FRAP assay, it was evaluated that LBTI had 87.5% and CCTI showed 84.4% antioxidant activity, taking value of standard ascorbic acid to be 100%. The PPIs also showed in-vitro anti‐inflammatory activity by inhibiting the heat induced albumin denaturation with IC50 values of 686 µg/ml and 615 µg/ml for CCTI and LBTI respectively compared to the standard (aspirin) which was 70.8 µg/ml. Red blood cells membrane stabilization with IC50 values of 641 µg/ml and 587 µg/ml for CCTI and LBTI respectively against aspirin which showed IC50 value of 70.4 µg/ml. PPIs showed antibacterial activity against 7 known strains while there was apparently no action against fungi.

Keywords: Cajanus cajan, Phaseolus limensis, Lima beans, protein protease inhibitor, antioxidant, anti-inflammatory, antimicrobial activity

Procedia PDF Downloads 283

Authors: Mohammad Shamim Hasan Mandal, Phu Minh, Do Tan Khang, Phung Thi Tuyen, Tran Dang Xuan

Abstract:

Rice (Oryza sativa L.) is one of the major crops of Vietnam which has more than thousands of varieties. Many of the local varieties have greater potentiality but they are in danger of extinct. Rice plant contains many secondary metabolites that are allelopathic to other plants. Seven rice varieties were cultivated in the field condition at Hiroshima University, Japan; stems and leaves from each variety were collected later, they were extracted with methanol, hexane, ethyl acetate, butanol, and water. Total phenolic content and total flavonoid contents were high in ethyl acetate extracts. DPPH antioxidant assay results showed that the ethyl acetate extracts had the higher IC50 value. Therefore, the ethyl acetate extracts were selected for laboratory experimentation through petri dish assay. Results showed that the two-local variety Re nuoc and Nan chon completely inhibited the germination of radish seedlings. Further laboratory bioassay and field experimentation will be conducted to validate the laboratory bioassay findings.

Keywords: allelopathy, bioassay, Oryza sativa, Raphanus sativus

Procedia PDF Downloads 348

Authors: Jacky Bhagat, Baban S Ingole

Abstract:

In this study, in vivo experiments were carried out to investigate the effects of a toxic polycyclic aromatic hydrocarbon (PAH), benzo(k)fluoranthene (B[k]F), on marine gastropod, Morula granulata collected from Goa, west coast of India. Snails were exposed to different concentrations of B(k)F (1, 10, 25 and 50 µg/L) for 96 h. The genotoxic effects were evaluated by measuring DNA strand breaks using alkaline comet assay and oxidative stress were measured with the help of battery of biomarkers such as superoxide dismutase (SOD) catalase (CAT), glutathione-s-transferase (GST), and lipid peroxidation (LPO). Concentration-dependent increase in percentage tail DNA (TDNA) was observed in snails exposed to B(k)F. Exposure concentrations above 1 µg/L of B(k)F, showed significant increase in SOD activity and LPO value in snails. After 96 h, SOD activity were found to be doubled for 50 µg/L of B(k)F with reference to control. Significant increase in CAT and GST activity was observed at all exposure conditions at the end of the exposure time. Our study showed that B(k)F induces oxidative stress in snails which further lead to genotoxic damage.

Keywords: benzo(k)fluoranthene, comet assay, gastropod, oxidative stress

Procedia PDF Downloads 325

Authors: Hammad Khan, Wookeun Bae

Abstract:

The objective of this study was to optimize and control the highly loaded and efficient nitrite production having suitability for autotrophic and heterotrophic denitritation. A lab scale CSTR for partial and full nitritation was operated to treat the livestock manure digester liquor having an ammonium concentration of ~600 mg-NH4+-N/L and biodegradable contents of ~0.35 g-COD/L. The experiments were performed at 30°C, pH: 8.0, DO: 1.5 mg/L and SRT ranging from 7-20 days. After 125 days operation, >95% nitrite buildup having the ammonium loading rate of ~3.2 kg-NH4+-N/m3-day was seen with almost complete ammonium conversion. On increasing the loading rate further (i-e, from 3.2-6.2 kg-NH4+-N/m3-day), stability of the system remained unaffected. On decreasing the pH from 8 to 7.5 and further 7.2, removal rate can be easily controlled as 95%, 75%, and even 50%. Results demonstrated that nitritation stability and desired removal rates are controlled by a balance of simultaneous inhibition by FA & FNA, pH effect and DO limitation. These parameters proved to be effective even to produce an appropriate influent for anammox. In addition, a mathematical model, identified through the occurring biological reactions, is proposed to optimize the full and partial nitritation process. The proposed model present relationship between pH, ammonium and produced nitrite for full and partial nitritation under the varying concentrations of DO, and simultaneous inhibition by FA and FNA.

Keywords: stable nitritation, high loading, autrophic denitritation, hetrotrophic denitritation

Procedia PDF Downloads 297

Authors: Hammad Khan, Wookeun Bae

Abstract:

The objective of this study was to optimize, achieve and control the highly loaded and efficient nitrite production having suitability for autotrophic and heterotrophic denitritation. A lab scale CSTR for partial and full nitritation was operated to treat the livestock manure digester liquor having an ammonium concentration of ~600 mg-NH4+-N/L and biodegradable contents of ~0.35 g-COD/L. The experiments were performed at 30°C, pH: 8.0, DO: 1.5 mg/L and SRT ranging from 7-20 days. After 125 days operation, >95% nitrite buildup having the ammonium loading rate of ~3.2 kg-NH4+-N/m3-day was seen with almost complete ammonium conversion. On increasing the loading rate further (i-e, from 3.2-6.2 kg-NH4+-N/m3-day), stability of the system remained unaffected. On decreasing the pH from 8 to7.5 and further 7.2, removal rate can be easily controlled as 95%, 75%, and even 50%. Results demonstrated that nitritation stability and desired removal rates are controlled by a balance of simultaneous inhibition by FA & FNA, pH affect and DO limitation. These parameters proved to be effective even to produce an appropriate influent for anammox. In addition, a mathematical model, identified through the occurring biological reactions, is proposed to optimize the full and partial nitritation process. The proposed model present relationship between pH, ammonium and produced nitrite for full and partial nitritation under the varying concentrations of DO, and simultaneous inhibition by FA and FNA.

Keywords: stable nitritation, high loading, autrophic denitritation, CSTR

Procedia PDF Downloads 222

Authors: Po-Sheng Kuo, Che-Wei Lu, Ssu-Ching Chen

Abstract:

Chlorinated ethenes (CEs) constitute a predominant contaminant in Taiwan's native polluted sites, particularly in groundwater inundated with sulfate salts that substantially impede remediation efforts. The reduction of sulfate by sulfate-reducing bacteria (SRB) impairs the dechlorination efficiency of Dehalococcoides by generating hydrogen sulfide (H₂S), resulting in incomplete chloride degradation and thereby leading to the failure of bioremediation. In order to elucidate interactions between sulfate reduction and dechlorination, this study aims to establish a co-culture system of Dehalococcoides and SRB, overcoming H₂S inhibition by employing the synthesis of ferrous sulfide (FeS), which is commonly utilized in chemical remediation due to its high reduction potential. Initially, the study demonstrates that the addition of ferrous chloride (FeCl₂) effectively removed H₂S production from SRB and enhanced the degradation of trichloroethylene to ethene. This process overcomes the inhibition caused by H₂S produced by SRB in high sulfate environments. Compared to different concentrations of ferrous dosages for the biogenic generation of FeS, the efficiency was optimized by adding FeCl₂ at an equal ratio to the concentration of sulfate in the environment. This was more effective in removing H₂S and crystal particles under 10 times smaller than those synthesized under excessive FeCl₂ dosages, addressing clogging issues in situ remediation. Finally, utilizing Taiwan's indigenous dechlorinating consortium in a simulated high sulfate-contaminated environment, the biodiversity of microbial species was analyzed to reveal a higher species richness within the FeS group, conducive to ecological stability. This study validates the potential of the co-culture system in generating biogenic FeS under sulfate and CEs co-contamination, removing sulfate-reducing products, and improving CE remediation through integrated chemical and biological remediations.

Keywords: biogenic ferrous sulfide, chlorinated ethenes, Dehalococcoides, sulfate-reducing bacteria, sulfide inhibition

Procedia PDF Downloads 41

Authors: Mehran Fadaeinasab, Hamed Karimian, Najihah Mohd Hashim, Hapipah Mohd Ali

Abstract:

In this study, three indole alkaloids namely; rauvolfine B, macusine B, and isoreserpiline have been isolated from the dichloromethane crude extract of Rauvolfia reflexa bark (Apocynaceae). The structural elucidation of the isolated compounds has been performed using spectral methods such as UV, IR, MS, 1D, and 2D NMR. Rauvolfine B showed anti proliferation activity on HCT-116 cancer cell line, its cytotoxicity induction was observed using MTT assay in eight different cell lines. Annexin-V is serving as a marker for apoptotic cells and the Annexin-V-FITC assay was carried out to observe the detection of cell-surface Phosphatidylserine (PS). Apoptosis was confirmed by using caspase-8 and -9 assays. Cell cycle arrest was also investigated using flowcytometric analysis. rauvolfine B had exhibited significantly higher cytotoxicity against HCT-116 cell line. The treatment significantly arrested HCT-116 cells in the S phase. Together, the results presented in this study demonstrated that rauvolfine B inhibited the proliferation of HCT-116 cells and programmed cell death followed by cell cycle arrest.

Keywords: apocynacea, indole alkaloid, apoptosis, cell cycle arrest

Procedia PDF Downloads 323

Authors: Sehrish Iftikhar, Ahmad Ali Shahid, Kiran Nawaz, Waheed Anwar

Abstract:

Discovery and development of new compounds require intense studies in chemistry, biochemistry. Numerous experiments under laboratory-, greenhouse- and field conditions can be performed to select suitable candidates and to understand their full potential. Novel fungicides are fundamental to combat plant diseases. Fusarium solani is important plant pathogen. New broad spectrum foliar fungicides against complex II were designed in this study. Complex II, namely succinate dehydrogenase (SDH), or succinate quinone oxidoreductase (SQR) is a multi-subunit enzyme at the crossroads of TCA and ETC at the inner mitochondrial membrane. The need for new and innovative fungicides is driven by resistance management, regulatory hurdles and increasing customer expectations amongst others. Fungicidal activity was assessed for the effect on mycelial growth and spore germination of the fungi using fungicide amended media assay. In mycelial growth assay compounds C10 and C6 were highly active against all the isolates. The compounds C1 and C10 were found most potent in spore germination test. It fully proved that the SDHIs designed in this paper displayed as good inhibitory effects against Fusarium solani.

Keywords: Wilt, Fusarium, SDH, antifungal

Procedia PDF Downloads 245

Authors: J. Carlos Kuri, Varun Vij, Raymond J. Turner, Orly Yadid-Pecht

Abstract:

Celiac disease (CD) is an immune system disorder that is triggered by ingesting gluten. As a gluten-free (GF) diet has become a concern of many people for health reasons, a gold standard had to be nominated. Enzyme-linked immunosorbent assay (ELISA) has taken the seat of this role. However, multiple limitations were discovered, and with that, the desire for an alternative method now exists. Nucleic acid-based aptamers have become of great interest due to their selectivity, specificity, simplicity, and rapid-testing advantages. However, fluorescence-based aptasensors have been tagged as unstable, but lifespan details are rarely stated. In this work, the lifespan stability of a fluorescence-based aptasensor is shown over an 8-week-long study displaying the accuracy of the sensor and false negatives. This study follows 22 different samples, including GF and gluten-rich (GR) and soy sauce products, off-the-shelf products, and reference material from laboratories, giving a total of 836 tests. The analysis shows an accuracy of correctly classifying GF and GR products of 96.30% and 100%, respectively when the protocol is augmented with molecular sieves. The overall accuracy remains around 94% within the first four weeks and then decays to 63%.

Keywords: aptasensor, PEG, rGO, FAM, RM, ELISA

Procedia PDF Downloads 115

Authors: Mahshid Hodjat, Maryam Baeeri, Mohammad Amin Rezvanfar, Mohammad Abdollahi

Abstract:

Ethephon is one of the most widely used plant growth regulator in agriculture that its application has been increased in recent years. The toxicity of organophosphate compounds is mostly attributed to their potent inhibition of acetylcholinesterase and their involvement in neurodegenerative disease. Although there are few reports on butyrylcholinesterase inhibitory role of ethephon, still there is no evidence on neurotoxicity and genotoxicity of this compound. The aim of the current study is to assess the potential genotoxic effect of ethephon using two genotoxic endpoints; γH2AX expression and comet assay on embryonic murine fibroblast. γH2AX serves as an early and sensitive biomarker for evaluating the genotoxic effects of chemicals. Oxidative stress biomarkers, including intracellular reactive oxygen species, lipid peroxidation and antioxidant capacity were also examined. The results showed a significant increase in cell proliferation 24h post-treatment with 10, 40,160µg/ml ethephon. The γH2AX expression and γH2AX foci count per cell were increased at low concentration of ethephon that was concomitant with increased DNA damage break at 40 and 160 µg/ml as illustrated by increased comet tail moment. A significant increase in lipid peroxidation and ROS formation were observed at 160 µg/ml and higher doses. The results showed that low-dose of ethephon promoted cell proliferation while induce DNA damage, raising the possibility of ethephon mutagenicity. Ethephon-induced genotoxic effect of low dose might not related to oxidative damage. However, ethephon was found to increase oxidative stress at higher doses, lead to cellular cytotoxicity. Taken together, all data indicated that ethylene, deserves more attention as a plant regulator with potential genotoxicity for which appropriate control is needed to reduce its usage.

Keywords: ethephon, DNA damage, γH2AX, oxidative stress

Procedia PDF Downloads 297

Authors: Constance Chivengwa, Tinashe Mandimutsira, Jephris Gere, Charles Magogo, Irene Chikanza, Jerneja Vidmar, Walter Chingwaru

Abstract:

The use of traditional methods in the management of diarrhoea has remained a common practice among the indigenous African tribes of Southern Africa. Despite the widespread use of traditional medicines in Zimbabwe, very little research validating the activities of phytomedicines against diarrhoea, as claimed by the Shona people of Zimbabwe, has been reported. This study sought to determine the efficacies of the plants that are frequently used to treat stomach complaints, namely Dicoma anomala, Cassia abbreviata, Lannea edulis and Peltophorum africanum against Escherichia coli (an indicator of faecal contamination of water, and whose strains such as EHEC (O157), ETEC and EPEC, are responsible for a number of outbreaks of diarrhoea) and Salmonella spp. Ethanol and aqueous extracts from these plants were obtained, evaporated, dried and stored. The dried extracts were reconstituted and diluted 10-fold in nutrient broth (from 100 to 0.1 microgram/mL) and tested for inhibition against the bacteria. L. edulis exhibited the best antimicrobial effect (minimum inhibition concentration = 10 microgram/mL for both extracts and microorganisms). Runners up to L. edulis were C. abbreviata (20 microgram/mL for both microorganisms) and P. africanum (20 and 30 microgram/mL respectively). Interestingly, D. anomala, which is widely considered panacea in African medicinal practices, showed low antimicrobial activity (60 and 100 microgram/mL respectively). The high antimicrobial activity of L. edulis can be explained by its content of flavonoids, tannins, alkylphenols (cardonol 7 and cardonol 13) and dihydroalkylhexenones. The antimicrobial activities of C. abbreviata can be linked to its content of anthraquinones and triterpenoids. P. africanum is known to contain benzenoids, flavanols, flavonols, terpenes, xanthone and coumarins. This study therefore demonstrated that, among the plants that are used against diarrhoea in African traditional medicine, L. edulis is a clear winner against E. coli and Salmonella spp. Activity guided extraction is encouraged to establish the complement of compounds that have antimicrobial activities.

Keywords: diarrhoea, Escherichia coli, Salmonella, phytomedicine, MIC, Zimbabwe

Procedia PDF Downloads 355

Authors: Assia Belfar, Mohamed Hadjadj, Messaouda Dakmouche, Zineb Ghiaba, Mahdi Belguidoum

Abstract:

Introduction: This study aims to phytochemical screening; Extracting the active compounds and estimate the effectiveness of antioxidant in Medicinal plants desert Limoniastrum guyonianum (Zeïta) from South Algeria. Methods: Total phenolic content and total flavonoid content using Folin-Ciocalteu and aluminum chloride colorimetric methods, respectively. The total antioxidant capacity was estimated by the following methods: DPPH (1.1-diphenyl-2-picrylhydrazyl radical) and reducing power assay. Results: Phytochemical screening of the plant part reveals the presence of phenols, saponins, flavonoids and tannins. While alkaloids and Terpenoids were absent. The Methanolic extract of L. guyonianum was extracted successively with ethyl acetate and butanol. Extraction of yield varied widely in the L. guyonianum ranging from (1.315 % to 4.218%). butanol fraction had the highest yield. The higher content of phenols was recorded in butanol fraction (311.81 ± 0.02mg GAE/g DW), the higher content of flavonoids was found in butanol fraction (9.58 ± 0.33mg QE/g DW). IC50 of inhibition of radical DPPH in ethyl acetate fraction was (0.05 ± 0.01µg/ml) Equal effectiveness with BHT, All extracts showed good activity of ferric reducing power, the higher power was in butanol fraction (16.16 ± 0.05mM). Conclusions: Demonstrated this study that the Methanolic extract of L. guyonianum contain a considerable quantity of phenolic compounds and possess a good antioxidant activity. It can be used as an easily accessible source of Natural Antioxidants and as a possible food supplement and in pharmaceutical industry.

Keywords: flavonoid compound, l. guyonianum, medicinal plants, phenolic compounds, phytochemical screening

Procedia PDF Downloads 289

Authors: Ronald Croce, Timothy Quinn, John Miller

Abstract:

Incomplete activation, or activation failure, of motor units during maximal voluntary contractions is often referred to as muscle inhibition (MI), and is defined as the inability of the central nervous system to maximally drive a muscle during a voluntary contraction. The purpose of the present study was to assess the interrelationship amongst peak torque (PT), muscle inhibition (MI; incomplete activation of motor units), and voluntary muscle activation (VMA) of the quadriceps’ muscle group as a function of knee angle and muscle length during maximal voluntary isometric contractions (MVICs). Nine young adult males (mean + standard deviation: age: 21.58 + 1.30 years; height: 180.07 + 4.99 cm; weight: 89.07 + 7.55 kg) performed MVICs in random order with the knee at 15, 55, and 95° flexion. MI was assessed using the interpolated twitch technique and was estimated by the amount of additional knee extensor PT evoked by the superimposed twitch during MVICs. Voluntary muscle activation was estimated by root mean square amplitude electromyography (EMGrms) and mechanomyography (MMGrms) of agonist (vastus medialis [VM], vastus lateralis [VL], and rectus femoris [RF]) and antagonist (biceps femoris ([BF]) muscles during MVICs. Data were analyzed using separate repeated measures analysis of variance. Results revealed a strong dependency of quadriceps’ PT (p < 0.001), MI (p < 0.001) and MA (p < 0.01) on knee joint position: PT was smallest at the most shortened muscle position (15°) and greatest at mid-position (55°); MI and MA were smallest at the most shortened muscle position (15°) and greatest at the most lengthened position (95°), with the RF showing the greatest change in MA. It is hypothesized that the ability to more fully activate the quadriceps at short compared to longer muscle lengths (96% contracted at 15°; 91% at 55°; 90% at 95°) might partly compensate for the unfavorable force-length mechanics at the more extended position and consequent declines in VMA (decreases in EMGrms and MMGrms muscle amplitude during MVICs) and force production (PT = 111-Nm at 15°, 217-NM at 55°, 199-Nm at 95°). Biceps femoris EMG and MMG data showed no statistical differences (p = 0.11 and 0.12, respectively) at joint angles tested, although there were greater values at the extended position. Increased BF muscle amplitude at this position could be a mechanism by which anterior shear and tibial rotation induced by high quadriceps’ activity are countered. Measuring and understanding the degree to which one sees MI and VMA in the QF muscle has particular clinical relevance because different knee-joint disorders, such ligament injuries or osteoarthritis, increase levels of MI observed and markedly reduced the capability of full VMA.

Keywords: electromyography, interpolated twitch technique, mechanomyography, muscle activation, muscle inhibition

Procedia PDF Downloads 333

Authors: Azimeh Rajaee, Lingyun Zhao, Shi Wang, Yaqiang Liu

Abstract:

In recent years many studies have been focused on bismuth-based nanoparticles as radiosensitizer and contrast agent in radiation therapy and imaging due to the high atomic number (Z = 82), high photoelectric absorption, low cost, and low toxicity. This study aims to introduce a new multifunctional bismuth-based nanoparticle as a theranostic agent for radiotherapy, computed tomography (CT) and magnetic resonance imaging (MRI). We synthesized bismuth ferrite (BFO, BiFeO3) nanoparticles by sol-gel method and surface of the nanoparticles were modified by Polyethylene glycol (PEG). After proved biocompatibility of the nanoparticles, the ability of them as contract agent in Computed tomography (CT) and magnetic resonance imaging (MRI) was investigated. The relaxation time rate (R2) in MRI and Hounsfield unit (HU) in CT imaging were increased with the concentration of the nanoparticles. Moreover, the effect of nanoparticles on dose enhancement in low energy was investigated by clonogenic assay. According to clonogenic assay, sensitizer enhancement ratios (SERs) were obtained as 1.35 and 1.76 for nanoparticle concentrations of 0.05 mg/ml and 0.1 mg/ml, respectively. In conclusion, our experimental results demonstrate that the multifunctional nanoparticles have the ability to employ as multimodal imaging and therapy to enhance theranostic efficacy.

Keywords: molecular imaging, nanomedicine, radiotherapy, theranostics

Procedia PDF Downloads 299

Authors: Hammad Khan, Wookeun Bae

Abstract:

The objective of this study was to optimize and control the highly loaded and efficient nitrite production having suitability for autotrophic and heterotrophic denitritation. A lab scale CSTR for partial and full nitritation was operated to treat the livestock manure digester liquor having an ammonium concentration of ~2000 mg-NH4+-N/L and biodegradable contents of ~0.8 g-COD/L. The experiments were performed at 30°C, pH: 8.0 DO: 1.5 mg/L and SRT ranging from 7-20 days. After 125 days operation, >95% nitrite buildup having the ammonium loading rate of ~3.2 kg-NH4+-N/m3-day was seen with almost complete ammonium conversion. On increasing the loading rate further (i.e. from 3.2-6.2 kg-NH4+-N/m3-day), stability of the system remained unaffected. On decreasing the pH from 8 to7.5 and further 7.2, removal rate can be easily controlled as 95%, 75% and even 50%. Results demonstrated that nitritation stability and desired removal rates are controlled by a balance of simultaneous inhibition by FA and FNA, pH affect and DO limitation. These parameters proved to be effective even to produce an appropriate influent for anammox. In addition, a mathematical model, identified through the occurring biological reactions, is proposed to optimize the full and partial nitritation process. The proposed model presents relationship between pH, ammonium and produced nitrite for full and partial nitritation under the varying concentrations of DO, and simultaneous inhibition by FA and FNA.

Keywords: stable nitritation, high loading, autrophic denitritation, hetrotrophic denitritation

Procedia PDF Downloads 311

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: Eddy Neuhaus, Hanif Shirinzadeh, Cigdem Karaaslan, Elif Ince, Hande Gurer-Orhan, Sibel Suzen

Abstract:

Reactive oxygen species (ROS) and oxidative stress can cause fatal damage to essential cell structures, including DNA. It is known that use of antioxidants could be advantageous in the prevention of various diseases such as cancer, cardiovascular diseases and neurodegenerative disorders. Since antioxidant properties of the indole ring-containing melatonin (MLT) has been described and evaluated, MLT-related compounds such as MLT metabolites and synthetic analogues are under investigation to determine which exhibit the highest activity with the lowest side-effects. Owing to indole and hydrazones appealing physiological properties and are mostly found in numerous biologically active compounds a series of indole-7-carbaldehyde hydrazone derivatives were synthesized, characterized and in vitro antioxidant activity was investigated by evaluating their reducing effect against oxidation of a redox-sensitive fluorescent probe. Cytotoxicity potential of all indole-based MLT analogues was investigated both by lactate dehydrogenase leakage assay and by MTT assay. This work was supported by The Scientific and Technological Research Council of Turkey (TÜBİTAK) Research and Development Grant 112S599.

Keywords: melatonin, antioxidant activity, indole, hydrazone, oxidative stress

Procedia PDF Downloads 473

Authors: S. Ibrahim, U. B. Abubakar, S. Danbirni, A. Usman, F. M. Ballah, C. A. Kudi, L. Lawson, G. H. Abdulrazak, I. A. Abdulkadir

Abstract:

Performing culture and characterization of mycobacteria in low resource settings like Nigeria is a very difficult task to undertake because of the very few and limited laboratories carrying out such an experiment; this is a largely due to stringent and laborious nature of the tests. Hence, a rapid, simple and accurate test for characterization is needed. The “SD BIOLINE TB Ag MPT 64 Rapid ®” is a simple and rapid immunochromatographic test used in differentiating Mycobacteria into Mycobacterium tuberculosis (NTM). The 100 sputa were obtained from patients suspected to be infected with tuberculosis and presented themselves to hospitals for check-up and treatment were involved in the study. The samples were cultured in a class III Biosafety cabinet and level III biosafety practices were followed. Forty isolates were obtained from the cultured sputa, and there were identified as Acid-fast bacilli (AFB) using Zeihl-Neelsen acid-fast stain. All the isolates (AFB positive) were then subjected to the SD BIOLINE Analyses. A total of 31 (77.5%) were characterized as MTBC, while nine (22.5%) were NTM. The total turnaround time for the rapid assay was just 30 minutes as compared to a few days of phenotypic and genotypic method. It was simple, rapid and reliable test to differentiate MTBC from NTM.

Keywords: culture, mycobacteria, non tuberculous mycobacterium, SD Bioline

Procedia PDF Downloads 328

Authors: Samah Lakehal, A. Meliani, F. Z. Benrebiha, C. Chaouia

Abstract:

In the last few years, due to the misuse of antibiotics and an increasing incidence of immunodeficiency-related diseases, the development of microbial drug resistance has become more and more of a pressing problem. Recently, natural products from medicinal plants represent a fertile ground for the development of novel antibacterial agents. Plants essential oils have come more into the focus of phytomedicine. The present study describes antimicrobial activity of Rosmarinus officinalis L. essential oil known medicinally for its powerful antibacterial properties. The essential oil of rosemary obtained by hydrodistillation (using Clevenger type apparatus) growing in Algeria (Djelfa city of south Algeria) was investigated by GC-MS. The essential oil yield of the study was 1.4 %. The major components were found to be camphor, camphene, 1,8-cineole. The essential oil has been tested for antimicrobial activity against eight bacteria (Gram-negative and Gram-positive), and three fungi including Candida albicans. Inhibition of growth was tested by the agar diffusion method based on the determination of the diameter of inhibition. The oil was found to have significant antibacterial activity and therefore can be used as a natural antimicrobial agent for the treatment of several infectious diseases caused by those germs, which have developed resistance to antibiotics.

Keywords: antimicrobial activity, Rosmarinus officinalis L., essential oils, GC/MS, camphor

Procedia PDF Downloads 381

Authors: Asma Ben Hmidene, Mizuho Hanaki, Kazuma Murakami, Kazuhiro Irie, Hiroko Isoda, Hideyuki Shigemori

Abstract:

Type 2 diabetes mellitus (T2DM) and Alzheimer’s disease (AD) are characterized as a peripheral metabolic disorder and a degenerative disease of the central nervous system, respectively. It is now widely recognized that T2DM and AD share many pathophysiological features including glucose metabolism, increased oxidative stress and amyloid aggregation. Amyloid beta (Aβ) is the components of the amyloid deposits in the AD brain and while the component of the amyloidogenic peptide deposit in the pancreatic islets of Langerhans is identified as human islet amyloid polypeptide (hIAPP). These two proteins are originated from the amyloid precursor protein and have a high sequence similarity. Although the amino acid sequences of amyloidogenic proteins are diverse, they all adopt a similar structure in aggregates called cross-beta-spine. Add at that, extensive studies in the past years have found that like Aβ1-42, IAPP forms early intermediate assemblies as spherical oligomers, implicating that these oligomers possess a common folding pattern or conformation. These similarities can be used in the search for effective pharmacotherapy for DM, since potent therapeutic agents such as antioxidants with a catechol moiety, proved to inhibit Aβ aggregation, may play a key role in the inhibit the aggregation of hIAPP treatment of patients with DM. Tamarix gallica is one of the halophyte species having a powerful antioxidant system. Although it was traditionally used for the treatment of various liver metabolic disorders, there is no report about the use of this plant for the treatment or prevention of T2DM and AD. Therefore, the aim of this work is to investigate their protective effect towards T2DM and AD by isolation and identification of α-glucosidase inhibitors, with antioxidant potential, that play an important role in the glucose metabolism in diabetic patient, as well as, the polymerization of hIAPP and Aβ aggregation inhibitors. Structure-activity relationship study was conducted for both assays. And as for α-glucosidase inhibitors, their mechanism of action and their synergistic potential when applied with a very low concentration of acarbose were also suggesting that they can be used not only as α-glucosidase inhibitors but also be combined with established α-glucosidase inhibitors to reduce their adverse effect. The antioxidant potential of the purified substances was evaluated by DPPH and SOD assays. Th-T assay using 42-mer amyloid β-protein (Aβ42) for AD and hIAPP which is a 37-residue peptide secreted by the pancreatic β –cells for T2DM and Transmission electronic microscopy (TEM) were conducted to evaluate the amyloid aggragation of the actives substances. For α-glucosidase, p-NPG and glucose oxidase assays were performed for determining the inhibition potential and structure-activity relationship study. The Enzyme kinetic protocol was used to study the mechanism of action. From this research, it was concluded that polyphenols playing a role in the glucose metabolism and oxidative stress can also inhibit the amyloid aggregation, and that substances with a catechol and glucuronide moieties inhibiting amyloid-β aggregation, might be used to inhibit the aggregation of hIAPP.

Keywords: α-glucosidase inhibitors, amyloid aggregation inhibition, mechanism of action, polyphenols, structure activity relationship, synergistic potential, tamarix gallica

Procedia PDF Downloads 268

Authors: Ibrahim Aly, Manal Ahmed, Mahmoud M. El-Shall

Abstract:

Filariasis is a parasitic disease caused by small roundworms. The filarial worms are transmitted and spread by blood-feeding black flies and mosquitoes. Lymphatic filariasis (Elephantiasis) is caused by Wuchereriabancrofti, Brugiamalayi, and Brugiatimori. Elimination of Lymphatic filariasis necessitates an increasing demand for valid, reliable, and rapid diagnostic kits. Nanodiagnostics involve the use of nanotechnology in clinical diagnosis to meet the demands for increased sensitivity, specificity, and early detection in less time. The aim of this study was to evaluate the nano-based enzymelinked immunosorbent assay (ELISA) and novel nano-chip card as a rapid test for detection of filarial antigen in serum samples of human filariasis in comparison with traditional -ELISA. Serum samples were collected from an infected human with filarial gathered across Egypt's governorates. After receiving informed consenta total of 45 blood samples of infected individuals residing in different villages in Gharbea governorate, which isa nonendemic region for bancroftianfilariasis, healthy persons living in nonendemic locations (20 persons), as well as sera from 20 other parasites, affected patients were collected. The microfilaria was checked in thick smears of 20 µl night blood samples collected during 20-22 hrs. All of these individuals underwent the following procedures: history taking, clinical examination, and laboratory investigations, which included examination of blood samples for microfilaria using thick blood film and serological tests for detection of the circulating filarial antigen using polyclonal antibody- ELISA, nano-based ELISA, and nano-chip card. In the present study, a recently reported polyoclonal antibody specific to tegumental filarial antigen was used in developing nano-chip card and nano-ELISA compared to traditional ELISA for the detection of circulating filarial antigen in sera of patients with bancroftianfilariasis. The performance of the ELISA was evaluated using 45 serum samples. The ELISA was positive with sera from microfilaremicbancroftianfilariasis patients (n = 36) with a sensitivity of 80 %. Circulating filarial antigen was detected in 39/45 patients who were positive for circulating filarial antigen using nano-ELISA with a sensitivity of 86.6 %. On the other hand, 42 out of 45 patients were positive for circulating filarial antigen using nano-chip card with a sensitivity of 93.3%.In conclusion, using a novel nano-chip assay could potentially be a promising alternative antigen detection test for bancroftianfilariasis.

Keywords: lymphatic filariasis, nanotechnology, rapid diagnosis, elisa technique

Procedia PDF Downloads 104