Search results for: beta lactamase inhibition
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1449

Search results for: beta lactamase inhibition

159 The Different Effects of Mindfulness-Based Relapse Prevention Group Therapy on QEEG Measures in Various Severity Substance Use Disorder Involuntary Clients

Authors: Yu-Chi Liao, Nai-Wen Guo, Chun‑Hung Lee, Yung-Chin Lu, Cheng-Hung Ko

Abstract:

Objective: The incidence of behavioral addictions, especially substance use disorders (SUDs), is gradually be taken seriously with various physical health problems. Mindfulness-based relapse prevention (MBRP) is a treatment option for promoting long-term health behavior change in recent years. MBRP is a structured protocol that integrates formal meditation practices with the cognitive-behavioral approach of relapse prevention treatment by teaching participants not to engage in reappraisal or savoring techniques. However, considering SUDs as a complex brain disease, questionnaires and symptom evaluation are not sufficient to evaluate the effect of MBRP. Neurophysiological biomarkers such as quantitative electroencephalogram (QEEG) may improve accurately represent the curative effects. This study attempted to find out the neurophysiological indicator of MBRP in various severity SUD involuntary clients. Participants and Methods: Thirteen participants (all males) completed 8-week mindfulness-based treatment provided by trained, licensed clinical psychologists. The behavioral data were from the Severity of Dependence Scale (SDS) and Negative Mood Regulation Scale (NMR) before and afterMBRP treatment. The QEEG data were simultaneously recorded with executive attention tasks, called comprehensive nonverbal attention test(CNAT). The two-way repeated-measures (treatment * severity) ANOVA and independent t-test were used for statistical analysis. Results: Thirteen participants regrouped into high substance dependence (HS) and low substance dependence (LS) by SDS cut-off. The HS group showed more SDS total score and lower gamma wave in the Go/No Go task of CNAT at pretest. Both groups showed the main effect that they had a lower frontal theta/beta ratio (TBR) during the simple reaction time task of CNAT. The main effect showed that the delay errors of CNAT were lower after MBRP. There was no other difference in CNAT between groups. However, after MBRP, compared to LS, the HS group have resonant progress in improving SDS and NMR scores. The neurophysiological index, the frontal TBR of the HS during the Go/No Go task of CNATdecreased than that of the LS group. Otherwise, the LS group’s gamma wave was a significant reduction on the Go/No Go task of CNAT. Conclusion: The QEEG data supports the MBRP can restore the prefrontal function of involuntary addicts and lower their errors in executive attention tasks. However, the improvement of MBRPfor the addict with high addiction severity is significantly more than that with low severity, including QEEG’s indicators and negative emotion regulation. Future directions include investigating the reasons for differences in efficacy among different severity of the addiction.

Keywords: mindfulness, involuntary clients, QEEG, emotion regulation

Procedia PDF Downloads 128
158 The Quantitative Optical Modulation of Dopamine Receptor-Mediated Endocytosis Using an Optogenetic System

Authors: Qiaoyue Kuang, Yang Li, Mizuki Endo, Takeaki Ozawa

Abstract:

G protein-coupled receptors (GPCR) are the largest family of receptor proteins that detect molecules outside the cell and activate cellular responses. Of the GPCRs, dopamine receptors, which recognize extracellular dopamine, are essential to mammals due to their roles in numerous physiological events, including autonomic movement, hormonal regulation, emotions, and the reward system in the brain. To precisely understand the physiological roles of dopamine receptors, it is important to spatiotemporally control the signaling mediated by dopamine receptors, which is strongly dependent on their surface expression. Conventionally, chemical-induced interactions were applied to trigger the endocytosis of cell surface receptors. However, these methods were subjected to diffusion and therefore lacked temporal and special precision. To further understand the receptor-mediated signaling and to control the plasma membrane expression of receptors, an optogenetic tool called E-fragment was developed. The C-terminus of a light-sensitive photosensory protein cyptochrome2 (CRY2) was attached to β-Arrestin, and the E-fragment was generated by fusing the C-terminal peptide of vasopressin receptor (V2R) to CRY2’s binding partner protein CIB. The CRY2-CIB heterodimerization triggered by blue light stimulation brings β-Arrestin to the vicinity of membrane receptors and results in receptor endocytosis. In this study, the E-fragment system was applied to dopamine receptors 1 and 2 (DRD1 and DRD2) to control dopamine signaling. First, confocal fluorescence microscope observation qualitatively confirmed the light-induced endocytosis of E-fragment fused receptors. Second, NanoBiT bioluminescence assay verified quantitatively that the surface amount of E-fragment labeled receptors decreased after light treatment. Finally, GloSensor bioluminescence assay results suggested that the E-fragment-dependent receptor light-induced endocytosis decreased cAMP production in DRD1 signaling and attenuated the inhibition effect of DRD2 on cAMP production. The developed optogenetic tool was able to induce receptor endocytosis by external light, providing opportunities to further understand numerous physiological activities by controlling receptor-mediated signaling spatiotemporally.

Keywords: dopamine receptors, endocytosis, G protein-coupled receptors, optogenetics

Procedia PDF Downloads 67
157 Search of Сompounds with Antimicrobial and Antifungal Activity in the Series of 1-(2-(1H-Tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas

Authors: O. Antypenko, I. Vasilieva, S. Kovalenko

Abstract:

Investigations for new effective and less toxic antimicrobials agents are always up-to-date. The tetrazole derivatives are quite interesting objects as for synthesis as well as for pharmacological screening. Thus, some derivatives of tetrazole demonstrated antimicrobial activity, namely 5-phenyl-tetrazolo[1,5-c]quinazoline was effective one against Staphylococcus aureus and Esherichia faecalis (MIC = 250 mg/L). Besides, investigation of the 9-bromo(chloro)-5-morpholin(piperidine)-4-yl-tetrazolo[1,5-c]quinazoline’s antimicrobial activity against Esherichia coli and Enterococcus faecalis, Pseudomonas aeruginosa and Staphylococcus aureus revealed that sensitivity of Gram-positive bacteria to the compounds was higher than that of Gram-negative bacteria. So, our previously synthesized, 31 derivatives of 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas were decided to test for their in vitro antibacterial activity against Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Enterobacter aerogenes, Enterococcus faecalis ATCC 29212), Gram-negative bacteria (Pseudomonas aeruginosa ATCC 9027, Escherichia coli ATCC 25922, Klebsiella pneumoniae 68) and antifungal properties against Candida albicans ATCC 885653. Agar-diffusion method was used for determination of the preliminary activity compared to well-known reference antimicrobials. All the compounds were dissolved in DMSO at a concentration of 100 μg/disk, using inhibition zone diameter (IZD, mm) as a measure for the antimicrobial activity. The most active turned to be 3 structures, that inhibited several bacterial strains: 1-ethyl-3-(5-fluoro-2-(1H-tetrazol-5-yl)phenyl)urea (1), 1-(4-bromo-2-(1H-tetrazol-5-yl)-phenyl)-3-(4-(trifluoromethyl)phenyl)urea (2) and 1-(4-chloro-2-(1H-tetrazol-5-yl)phenyl)-3-(3-(trifluoromethyl)phenyl)urea (3). IZM (mm) was 40 (Escherichia coli), 25 (Klebsiella pneumonia) for compound 1; 12 (Pseudomonas aeruginosa), 15 (Staphylococcus aureus), 10 (Enterococcus faecalis) for compound 2; 25 (Staphylococcus aureus), 15 (Enterococcus faecalis) for compound 3. The most sensitive to the activity of the substances were Gram-negative bacteria Pseudomonas aeruginosa. While none of compound effected on Candida albicans. Speaking about, reference drugs: Amikacin (30 µg/disk) showed 27 and Ceftazide (30 µg/disk) 25 against Pseudomonas aeruginosa. That is, unfortunately, higher than studied 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas. Obtained results will be used for further purposeful optimization of the leading compounds in the more effective antimicrobials because of the ever-mounting problem of microorganism’s resistance.

Keywords: antimicrobial, antifungal, compounds, 1-(2-(1H-tetrazol-5-yl)-R1-phenyl)-3-R2-phenyl(ethyl)ureas

Procedia PDF Downloads 336
156 Sustainable Production of Algae through Nutrient Recovery in the Biofuel Conversion Process

Authors: Bagnoud-Velásquez Mariluz, Damergi Eya, Grandjean Dominique, Frédéric Vogel, Ludwig Christian

Abstract:

The sustainability of algae to biofuel processes is seriously affected by the energy intensive production of fertilizers. Large amounts of nitrogen and phosphorus are required for a large-scale production resulting in many cases in a negative impact of the limited mineral resources. In order to meet the algal bioenergy opportunity it appears crucial the promotion of processes applying a nutrient recovery and/or making use of renewable sources including waste. Hydrothermal (HT) conversion is a promising and suitable technology for microalgae to generate biofuels. Besides the fact that water is used as a “green” reactant and solvent and that no biomass drying is required, the technology offers a great potential for nutrient recycling. This study evaluated the possibility to treat the water HT effluent by the growth of microalgae while producing renewable algal biomass. As already demonstrated in previous works by the authors, the HT aqueous product besides having N, P and other important nutrients, presents a small fraction of organic compounds rarely studied. Therefore, extracted heteroaromatic compounds in the HT effluent were the target of the present research; they were profiled using GC-MS and LC-MS-MS. The results indicate the presence of cyclic amides, piperazinediones, amines and their derivatives. The most prominent nitrogenous organic compounds (NOC’s) in the extracts were carefully examined by their effect on microalgae, namely 2-pyrrolidinone and β-phenylethylamine (β-PEA). These two substances were prepared at three different concentrations (10, 50 and 150 ppm). This toxicity bioassay used three different microalgae strains: Phaeodactylum tricornutum, Chlorella sorokiniana and Scenedesmus vacuolatus. The confirmed IC50 was for all cases ca. 75ppm. Experimental conditions were set up for the growth of microalgae in the aqueous phase by adjusting the nitrogen concentration (the key nutrient for algae) to fit that one established for a known commercial medium. The values of specific NOC’s were lowered at concentrations of 8.5 mg/L 2-pyrrolidinone; 1mg/L δ-valerolactam and 0.5 mg/L β-PEA. The growth with the diluted HT solution was kept constant with no inhibition evidence. An additional ongoing test is addressing the possibility to apply an integrated water cleanup step making use of the existent hydrothermal catalytic facility.

Keywords: hydrothermal process, microalgae, nitrogenous organic compounds, nutrient recovery, renewable biomass

Procedia PDF Downloads 386
155 Production of Functional Crackers Enriched with Olive (Olea europaea L.) Leaf Extract

Authors: Rosa Palmeri, Julieta I. Monteleone, Antonio C. Barbera, Carmelo Maucieri, Aldo Todaro, Virgilio Giannone, Giovanni Spagna

Abstract:

In recent years, considerable interest has been shown in the functional properties of foods, and a relevant role has been played by phenolic compounds, able to scavenge free radicals. A more sustainable agriculture has to emerge to guarantee food supply over the next years. Wheat, corn, and rice are the most common cereals cultivated, but also other cereal species, such as barley can be appreciated for their peculiarities. Barley (Hordeum vulgare L.) is a C3 winter cereal that shows high resistance at drought and salt stresses. There are growing interests in barley as ingredient for the production of functional foods due to its high content of phenolic compounds and Beta-glucans. In this respect, the possibility of separating specific functional fractions from food industry by-products looks very promising. Olive leaves represent a quantitatively significant by-product of olive grove farming, and are an interesting source of phenolic compounds. In particular, oleuropein, which provide important nutritional benefits, is the main phenolic compound in olive leaves and ranges from 17% to 23% depending upon the cultivar and growing season period. Together with oleuropein and its derivatives (e.g. dimethyloleuropein, oleuropein diglucoside), olive leaves further contain tyrosol, hydroxytyrosol, and a series of secondary metabolities structurally related to them: verbascoside, ligstroside, hydroxytyrosol glucoside, tyrosol glucoside, oleuroside, oleoside-11-methyl ester, and nuzhenide. Several flavonoids, flavonoid glycosides, and phenolic acids have also described in olive leaves. The aim of this work was the production of functional food with higher content of polyphenols and the evaluation of their shelf life. Organic durum wheat and barley grains contain higher levels of phenolic compounds were used for the production of crackers. Olive leaf extract (OLE) was obtained from cv. ‘Biancolilla’ by aqueous extraction method. Two baked goods trials were performed with both organic durum wheat and barley flours, adding olive leaf extract. Control crackers, made as comparison, were produced with the same formulation replacing OLE with water. Total phenolic compound, moisture content, activity water, and textural properties at different time of storage were determined to evaluate the shelf-life of the products. Our the preliminary results showed that the enriched crackers showed higher phenolic content and antioxidant activity than control. Alternative uses of olive leaf extracts for crackers production could represent a good candidate for the addition of functional ingredients because bakery items are daily consumed, and have long shelf-life.

Keywords: barley, functional foods, olive leaf, polyphenols, shelf life

Procedia PDF Downloads 277
154 The Effect of SIAH1 on PINK1 Homeostasis in Parkinson Disease

Authors: Fatimah Abd Elghani, Raymonde Szargel, Vered Shani, Hazem Safory, Haya Hamza, Mor Savyon, Ruth Rott, Rina Bandopadhyay, Simone Engelender

Abstract:

Background: PINK1 is a mitochondrial kinase mutated in some familial cases of Parkinson’s disease. Down regulation of PINK1 results in abnormal mitochondrial morphology and altered membrane potential. Although PINK1 has a predicted mitochondrial import sequence, it’s cellular, and submitochondrial localization remains unclear, in part because it is rapidly degraded. In this work, we investigated the mechanisms involved in PINK1 degradation and how this may affect PINK1 stability and function, with implications for mitochondrial function in PD. In addition, pharmacological inhibition of proteasome activity was shown to lead to PINK1 accumulation, indicating that PINK1 degradation depends on the ubiquitin-proteasome system (UPS). Methods: Using co-immunoprecipitation assays, we identified E3 ubiquitin ligase SIAH1 as a PINK1-interacting protein in HEK293 cells as well as on rat brain tissues. In addition, we determined the effect of SIAH 1, SIAH2 and Parkin on PINK1 steady-state levels by Western blot analysis, and checked their possibility to ubiquitinate and mediate PINK1 degradation through the proteasome carried out in vivo ubiquitination experiments. Results: We have obtained results showing that SIAH-1 interacts with and ubiquitinates PINK1. The ubiquitination promoted by SIAH-1 leads to the proteasomal degradation of PINK1. We confirmed these findings by knocking down SIAH-1 and observing important accumulation of PINK1 in cells. Besides, we found that SIAH-1 decreases PINK1 steady-state levels but not the E3 ligase Parkin. We also investigated the interaction of SIAH-1 with PINK1 disease mutants and its ability to promote their ubiquitination and degradation. Although, no clear difference in the ability of SIAH-1 to promote the degradation of PINK1 disease mutants was observed. It is possible that dysfunction of proteasomal activity in the disease may lead to the accumulation and aggregation of ubiquitinated PINK1 in patients with PINK1 mutations, with possible implications to the pathogenesis of PD. Conclusions: Here, we demonstrated that SIAH-1 ubiquitinates and promotes the degradation of PINK1. In addition, SIAH-1 represents now a target that may help the improvement of mitophagy in PD. Further investigations needed to understand how mitophagy is regulated by PINK1-SIAH-1 axis to provide targets for future therapeutics.

Keywords: PD, Parkinson's disease, PINK1, PTEN-induced kinase1, SIAH, seven in absentia homolog, SN, substantia nigra

Procedia PDF Downloads 118
153 Correlation Between the Toxicity Grade of the Adverse Effects in the Course of the Immunotherapy of Lung Cancer and Efficiency of the Treatment in Anti-PD-L1 and Anti-PD-1 Drugs - Own Clinical Experience

Authors: Anna Rudzińska, Katarzyna Szklener, Pola Juchaniuk, Anna Rodzajweska, Katarzyna Machulska-Ciuraj, Monika Rychlik- Grabowska, Michał łOziński, Agnieszka Kolak-Bruks, SłAwomir Mańdziuk

Abstract:

Introduction: Immune checkpoint inhibition (ICI) belongs to the modern forms of anti-cancer treatment. Due to the constant development and continuous research in the field of ICI, many aspects of the treatment are yet to be discovered. One of the less researched aspects of ICI treatment is the influence of the adverse effects on the treatment success rate. It is suspected that adverse events in the course of the ICI treatment indicate a better response rate and correlate with longer progression-free- survival. Methodology: The research was conducted with the usage of the documentation of the Department of Clinical Oncology and Chemotherapy. Data of the patients with a lung cancer diagnosis who were treated between 2019-2022 and received ICI treatment were analyzed. Results: Out of over 133 patients whose data was analyzed, the vast majority were diagnosed with non-small cell lung cancer. The majority of the patients did not experience adverse effects. Most adverse effects reported were classified as grade 1 or grade 2 according to CTCAE classification. Most adverse effects involved skin, thyroid and liver toxicity. Statistical significance was found for the adverse effect incidence and overall survival (OS) and progression-free survival (PFS) (p=0,0263) and for the time of toxicity onset and OS and PFS (p<0,001). The number of toxicity sites was statistically significant for prolonged PFS (p=0.0315). The highest OS was noted in the group presenting grade 1 and grade 2 adverse effects. Conclusions: Obtained results confirm the existence of the prolonged OS and PFS in the adverse-effects-charged patients, mostly in the group presenting mild to intermediate (Grade 1 and Grade 2) adverse effects and late toxicity onset. Simultaneously our results suggest a correlation between treatment response rate and the toxicity grade of the adverse effects and the time of the toxicity onset. Similar results were obtained in several similar research conducted - with the proven tendency of better survival in mild and moderate toxicity; meanwhile, other studies in the area suggested an advantage in patients with any toxicity regardless of the grade. The contradictory results strongly suggest the need for further research on this topic, with a focus on additional factors influencing the course of the treatment.

Keywords: adverse effects, immunotherapy, lung cancer, PD-1/PD-L1 inhibitors

Procedia PDF Downloads 62
152 Stems of Prunus avium: An Unexplored By-product with Great Bioactive Potential

Authors: Luís R. Silva, Fábio Jesus, Catarina Bento, Ana C. Gonçalves

Abstract:

Over the last few years, the traditional medicine has gained ground at nutritional and pharmacological level. The natural products and their derivatives have great importance in several drugs used in modern therapeutics. Plant-based systems continue to play an essential role in primary healthcare. Additionally, the utilization of their plant parts, such as leaves, stems and flowers as nutraceutical and pharmaceutical products, can add a high value in the natural products market, not just by the nutritional value due to the significant levels of phytochemicals, but also by to the high benefit for the producers and manufacturers business. Stems of Prunus avium L. are a byproduct resulting from the processing of cherry, and have been consumed over the years as infusions and decoctions due to its bioactive properties, being used as sedative, diuretic and draining, to relief of renal stones, edema and hypertension. In this work, we prepared a hydroethanolic and infusion extracts from stems of P. avium collected in Fundão Region (Portugal), and evaluate the phenolic profile by LC/DAD, antioxidant capacity, α-glucosidase inhibitory activity and protection of human erythrocytes against oxidative damage. The LC-DAD analysis allowed to the identification of 19 phenolic compounds, catechin and 3-O-caffolquinic acid were the main ones. In a general way, hydroethanolic extract proved to be more active than infusion. This extract had the best antioxidant activity against DPPH• (IC50=22.37 ± 0.28 µg/mL) and superoxide radical (IC50=13.93 ± 0.30 µg/mL). Furthermore, it was the most active concerning inhibition of hemoglobin oxidation (IC50=13.73 ± 0.67 µg/mL), hemolysis (IC50=1.49 ± 0.18 µg/mL) and lipid peroxidation (IC50=26.20 ± 0.38 µg/mL) on human erythrocytes. On the other hand, infusion revealed to be more efficient towards α-glucosidase inhibitory activity (IC50=3.18 ± 0.23 µg/mL) and against nitric oxide radical (IC50=99.99 ± 1.89 µg/mL). The Sweet cherry sector is very important in Fundão Region (Portugal), and taking profit from the great wastes produced during processing of the cherry to produce added-value products, such as food supplements cannot be ignored. Our results demonstrate that P. avium stems possesses remarkable antioxidant and free radical scavenging properties. It is therefore, suggest, that P. avium stems can be used as a natural antioxidant with high potential to prevent or slow the progress of human diseases mediated by oxidative stress.

Keywords: stems, Prunus avium, phenolic compounds, biological potential

Procedia PDF Downloads 272
151 A 4-Month Low-carb Nutrition Intervention Study Aimed to Demonstrate the Significance of Addressing Insulin Resistance in 2 Subjects with Type-2 Diabetes for Better Management

Authors: Shashikant Iyengar, Jasmeet Kaur, Anup Singh, Arun Kumar, Ira Sahay

Abstract:

Insulin resistance (IR) is a condition that occurs when cells in the body become less responsive to insulin, leading to higher levels of both insulin and glucose in the blood. This condition is linked to metabolic syndromes, including diabetes. It is crucial to address IR promptly after diagnosis to prevent long-term complications associated with high insulin and high blood glucose. This four-month case study highlights the importance of treating the underlying condition to manage diabetes effectively. Insulin is essential for regulating blood sugar levels by facilitating the uptake of glucose into cells for energy or storage. In IR individuals, cells are less efficient at taking up glucose from the blood resulting in elevated blood glucose levels. As a result of IR, beta cells produce more insulin to make up for the body's inability to use insulin effectively. This leads to high insulin levels, a condition known as hyperinsulinemia, which further impairs glucose metabolism and can contribute to various chronic diseases. In addition to regulating blood glucose, insulin has anti-catabolic effects, preventing the breakdown of molecules in the body, such as inhibiting glycogen breakdown in the liver, inhibiting gluconeogenesis, and inhibiting lipolysis. If a person is insulin-sensitive or metabolically healthy, an optimal level of insulin prevents fat cells from releasing fat and promotes the storage of glucose and fat in the body. Thus optimal insulin levels are crucial for maintaining energy balance and plays a key role in metabolic processes. During the four-month study, researchers looked at the impact of a low-carb dietary (LCD) intervention on two male individuals (A & B) who had Type-2 diabetes. Althoughvneither of these individuals were obese, they were both slightly overweight and had abdominal fat deposits. Before the trial began, important markers such as fasting blood glucose (FBG), triglycerides (TG), high-density lipoprotein (HDL) cholesterol, and Hba1c were measured. These markers are essential in defining metabolic health, their individual values and variability are integral in deciphering metabolic health. The ratio of TG to HDL is used as a surrogate marker for IR. This ratio has a high correlation with the prevalence of metabolic syndrome and with IR itself. It is a convenient measure because it can be calculated from a standard lipid profile and does not require more complex tests. In this four-month trial, an improvement in insulin sensitivity was observed through the ratio of TG/HDL, which, in turn, improves fasting blood glucose levels and HbA1c. For subject A, HbA1c dropped from 13 to 6.28, and for subject B, it dropped from 9.4 to 5.7. During the trial, neither of the subjects were taking any diabetic medications. The significant improvements in their health markers, such as better glucose control, along with an increase in energy levels, demonstrate that incorporating LCD interventions can effectively manage diabetes.

Keywords: metabolic disorder, insulin resistance, type-2 diabetes, low-carb nutrition

Procedia PDF Downloads 10
150 Profiling of Bacterial Communities Present in Feces, Milk, and Blood of Lactating Cows Using 16S rRNA Metagenomic Sequencing

Authors: Khethiwe Mtshali, Zamantungwa T. H. Khumalo, Stanford Kwenda, Ismail Arshad, Oriel M. M. Thekisoe

Abstract:

Ecologically, the gut, mammary glands and bloodstream consist of distinct microbial communities of commensals, mutualists and pathogens, forming a complex ecosystem of niches. The by-products derived from these body sites i.e. faeces, milk and blood, respectively, have many uses in rural communities where they aid in the facilitation of day-to-day household activities and occasional rituals. Thus, although livestock rearing plays a vital role in the sustenance of the livelihoods of rural communities, it may serve as a potent reservoir of different pathogenic organisms that could have devastating health and economic implications. This study aimed to simultaneously explore the microbial profiles of corresponding faecal, milk and blood samples from lactating cows using 16S rRNA metagenomic sequencing. Bacterial communities were inferred through the Divisive Amplicon Denoising Algorithm 2 (DADA2) pipeline coupled with SILVA database v138. All downstream analyses were performed in R v3.6.1. Alpha-diversity metrics showed significant differences between faeces and blood, faeces and milk, but did not vary significantly between blood and milk (Kruskal-Wallis, P < 0.05). Beta-diversity metrics on Principal Coordinate Analysis (PCoA) and Non-Metric Dimensional Scaling (NMDS) clustered samples by type, suggesting that microbial communities of the studied niches are significantly different (PERMANOVA, P < 0.05). A number of taxa were significantly differentially abundant (DA) between groups based on the Wald test implemented in the DESeq2 package (Padj < 0.01). The majority of the DA taxa were significantly enriched in faeces than in milk and blood, except for the genus Anaplasma, which was significantly enriched in blood and was, in turn, the most abundant taxon overall. A total of 30 phyla, 74 classes, 156 orders, 243 families and 408 genera were obtained from the overall analysis. The most abundant phyla obtained between the three body sites were Firmicutes, Bacteroidota, and Proteobacteria. A total of 58 genus-level taxa were simultaneously detected between the sample groups, while bacterial signatures of at least 8 of these occurred concurrently in corresponding faeces, milk and blood samples from the same group of animals constituting a pool. The important taxa identified in this study could be categorized into four potentially pathogenic clusters: i) arthropod-borne; ii) food-borne and zoonotic; iii) mastitogenic and; iv) metritic and abortigenic. This study provides insight into the microbial composition of bovine faeces, milk, and blood and its extent of overlapping. It further highlights the potential risk of disease occurrence and transmission between the animals and the inhabitants of the sampled rural community, pertaining to their unsanitary practices associated with the use of cattle by-products.

Keywords: microbial profiling, 16S rRNA, NGS, feces, milk, blood, lactating cows, small-scale farmers

Procedia PDF Downloads 83
149 Benefits of The ALIAmide Palmitoyl-Glucosamine Co-Micronized with Curcumin for Osteoarthritis Pain: A Preclinical Study

Authors: Enrico Gugliandolo, Salvatore Cuzzocrea, Rosalia Crupi

Abstract:

Osteoarthritis (OA) is one of the most common chronic pain conditions in dogs and cats. OA pain is currently viewed as a mixed phenomenon involving both inflammatory and neuropathic mechanisms at the peripheral (joint) and central (spinal and supraspinal) levels. Oxidative stress has been implicated in OA pain. Although nonsteroidal anti-inflammatory drugs are commonly prescribed for OA pain, they should be used with caution in pets because of adverse effects in the long term and controversial efficacy on neuropathic pain. An unmet need remains for safe and effective long-term treatments for OA pain. Palmitoyl-glucosamine (PGA) is an analogue of the ALIAamide palmitoylethanolamide, i.e., a body’s own endocannabinoid-like compound playing a sentinel role in nociception. PGA, especially in the micronized formulation, was shown safe and effective in OA pain. The aim of this study was to investigate the effect of a co-micronized formulation of PGA with the natural antioxidant curcumin (PGA-cur) on OA pain. Ten Sprague-Dawley male rats were used for each treatment group. The University of Messina Review Board for the care and use of animals authorized the study. On day 0, rats were anesthetized (5.0% isoflurane in 100% O2) and received intra-articular injection of MIA (3 mg in 25 μl saline) in the right knee joint, with the left being injected an equal volume of saline. Starting the third day after MIA injection, treatments were administered orally three times per week for 21 days, at the following doses: PGA 20 mg/kg, curcumin 10 mg/kg, PGA-cur (2:1 ratio) 30 mg/kg. On day 0 and 3, 7, 14 and 21 days post-injection, mechanical allodynia was measured using a dynamic plantar Von Frey hair aesthesiometer and expressed as paw withdrawal threshold (PWT) and latency (PWL). Motor functional recovery of the rear limb was evaluated on the same time points by walking track analysis using the sciatic functional index. On day 21 post-MIA injection, the concentration of the following inflammatory and nociceptive mediators was measured in serum using commercial ELISA kits: tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1β), nerve growth factor (NGF) and matrix metalloproteinase-1-3-9 (MMP-1, MMP-3, MMP-9). The results were analyzed by ANOVA followed by Bonferroni post-hoc test for multiple comparisons. Micronized PGA reduced neuropathic pain, as shown by the significant higher PWT and PWL values compared to vehicle group (p < 0.0001 for all the evaluated time points). The effect of PGA-cur was superior at all time points (p < 0.005). PGA-cur restored motor function already on day 14 (p < 0.005), while micronized PGA was effective a week later (D21). MIA-induced increase in the serum levels of all the investigated mediators was inhibited by PGA-cur (p < 0.01). PGA was also effective, except on IL-1 and MMP-3. Curcumin alone was inactive in all the experiments at any time point. The encouraging results suggest that PGA-cur may represent a valuable option in OA pain management and warrant further confirmation in well-powered clinical trials.

Keywords: ALIAmides, curcumin, osteoarthritis, palmitoyl-glucosamine

Procedia PDF Downloads 84
148 Identification and Antibiotic Susceptibility of Bacteria Isolated from the Intestines of Slaughtered Goat and Cattle

Authors: Latifat Afolake Ogunfolabo, Hakeem Babafemi Ogunfolabo

Abstract:

The gastrointestinal tract is densely populated with micro-organism which closely and intensively interacts with the host and ingested feed. Food borne infections are some of the major international challenges that lead to high mortality and also, antimicrobial resistance, which has been classified as a serious threat by World Health Organization. Samples of slaughtered cattle and goats intestines were collected and standard culture methods were used for bacteria isolation and identification. Minimum inhibitory concentration of commonly used antibiotic using modification of the disk diffusion method was carried out on isolates. The samples cultured were all positive to Pseudomonas aeruginosa (95% and 90%), Escherichia coli (85%), Salmonella typhi (70% and 60%), Staphylococcus aureus (75%and 100%), Micrococcus luteus (55% and35%), Bacillus macerans (60% and 5%), Bacillus cereus (25% and 20%), Clostridium perfringens (20% and 5%), Micrococcus varians (20% and 5%), Bacillus subtilis (25% and 5%), Streptococcus faecalis (40% and 25%) and Streptococcus faecium (15% and 10%) in goat and cattle respectively. Also, Proteus mirabilis (40%), Micrococcus luteus (35%), Proteus vulgaris (30%), Klebsiella aerogenes(15%) were isolated from cattle. The total coliform (13.55 x10⁵cfu/gm ± 1.77) and (20.30 x10⁵cfu/gm ± 1.27) counts were significantly higher than the total bacteria count (8.3 x10⁵cfu/gm ± 1.41) and (16.60 x10⁵cfu/gm ±0.49) for goat and cattle respectively. Selected Bacteria count of isolates showed that Staphylococcus aureus had the highest significant value (6.9 x10⁵cfu/gm ± 0.57) and (16.80 x10⁵cfu/gm ± 0.57) Escherichia coli (4.60 x10⁵cfu/gm ± 0.42) and (7.05 x10⁵cfu/gm ± 0.64) while the lowest significant value was obtained in Salmonella/Shigella (1.7 x10⁵cfu/gm ± 0.00) and (1.5 x10⁵cfu/gm ± 0.00) for goat and cattle respectively. Susceptibility of bacteria isolated from slaughtered goat and cattle intestine to commonly used antibiotics showed that the highest statistical significant value for zone of inhibition for goat was obtained for Ciprofloxacin (30.00 ± 2.25, 23.75 ± 2.49, 17.17 ± 1.40) followed by Augmentin (28.33 ± 1.22, 21. 83 ± 2.44, 16.67 ± 1.49), Erythromycin (27.75 ±1.48, 20.25 ± 1.29, 16.67 ± 1.26) while the lowest values were obtained for Ofloxacin (27.17 ± 1.89, 21.42 ± 2.19, 16.83 ± 1.26) respectively and values obtained for cattle are Ciprofloxacin (30.64 ± 1.6, 25.79 ± 1.76, 8.07 ± 11.49) followed by Augmentin (28.29 ± 1.33, 22.64 ± 1.82, 17.43 ± 1.55) Ofloxacin (26.57 ± 2.02, 20.79 ± 2.75, 16.21 ± 1.19) while the lowest values were obtained for Erythromycin (26.64 ± 1.49, 20.29 ± 1.49, 16.29 ± 1.33) at different dilution factor (10⁻¹, 10⁻², 10⁻³) respectively. The isolates from goat and cattle were all susceptible to Augmentin at the three different dilution factors. Some goat isolates are intermediate to Ciprofloxacin and Erythromycin at 10⁻² and 10⁻³, while resistance to Ciprofloxacin at 10⁻³ dilution factor. Ciprofloxacin and Ofloxacin at the dilution factors of 10⁻³ and 10⁻¹ for some cattle isolate and resistance were observed for Ofloxacin and Erythromycin at dilution of 10⁻³. These results indicate the susceptibilities and the antimicrobial resistance to commonly used antibiotic.

Keywords: antibiotic susceptibility, bacteria, cattle, goat, identification

Procedia PDF Downloads 95
147 The Effects of Nano Zerovalent Iron (nZVI) and Magnesium Oxide Nanoparticles on Methane Production during Anaerobic Digestion of Waste Activated Sludge

Authors: Passkorn Khanthongthip, John T. Novak

Abstract:

Many studies have been reported that the nZVI and MgO NPs were often found in waste activated sludge (WAS). However, little is known about the impact of those NPs on WAS stabilization. The aims of this study were to investigate the effects of both NPs on WAS anaerobic digestion for methane production and to examine the change of metanogenic population under those different environments using qPCR. Four dosages (2, 50, 100, and 200 mg/g-TSS) of MgO NPs were added to four different bottles containing WAS to investigate the impact of MgO NPs on methane production during WAS anaerobic digestion. The effects of nZVI on methane production during WAS anaerobic digestion were also conducted in another four bottles using the same methods described above except that the MgO NPs were replaced by nZVI. A bottle of WAS anaerobic digestion without nanoparticles addition was also operated to serve as a control. It was found that the relative amounts, compared to the control system, of methane production in each WAS anaerobic digestion bottle adding 2, 50, 100, 200 mg/gTSS MgO NPs were 98, 62, 28, and 14 %, respectively. This suggests that higher MgO NPs resulted in lower methane production. The data of batch test for the effects of corresponding released Mg2+ indicated that 50 mg/gTSS MgO NPs or higher could inhibit methane production at least 25%. Moreover, the volatile fatty acid (VFA) concentration was 328, 384, 928, 3,684, and 7,848 mg/L for the control and four WAS anaerobic digestion bottles with 2, 50, 100, 200 mg/gTSS MgO NPs addition, respectively. Higher VFA concentration could reduce pH and subsequently decrease methanogen growth, resulting in lower methane production. The relative numbers of total gene copies of methanogens analyzed from samples taken from WAS anaerobic digestion bottles were approximately 99, 68, 38, and 24 % of control for the addition of 2, 50, 100, and 200 mg/gTSS, respectively. Obviously, the more MgO NPs appeared in sludge anaerobic digestion system, the less methanogens remained. In contrast, the relative amount of methane production found in another four WAS anaerobic digestion bottles adding 2, 50, 100, and 200 mg/gTSS nZVI were 102, 128, 112, and 104 % of the control, respectively. The measurement of methanogenic population indicated that the relative content of methanogen gene copies were 101, 132, 120, and 112 % of those found in control, respectively. Additionally, the cumulative VFA was 320, 234, 308, and 330 mg/L, respectively. This reveals that nZVI addition could assist to increase methanogenic population. Higher amount of methanogen accelerated VFA degradation for greater methane production, resulting in lower VFA accumulation in digesters. Moreover, the data for effects of corresponding released Fe2+ conducted by batch tests suggest that the addition of approximately 50 mg/gTSS nZVI increased methane production by 20%. In conclusion, the presence of MgO NPs appeared to diminish the methane production during WAS anaerobic digestion. Higher MgO NPs dosages resulted in more inhibition on methane production. In contrast, nZVI addition promoted the amount of methanogenic population which facilitated methane production.

Keywords: magnesium oxide nanoparticles, methane production, methanogenic population, nano zerovalent iron

Procedia PDF Downloads 273
146 Corrosion Behavior of Organic-Inorganic Hybrid Coatings Fabricated by Electrostatic Method

Authors: Mohammed Ahmed, Ziba Nazarlou

Abstract:

Mild steels have a limited alloying content which makes them vulnerable to excessive corrosion rates in the harsh medium. To overcome this issue, some protective coatings are used to prevent corrosion on the steel surface. The use of specialized coatings, mainly organic coatings (such as epoxies, polyurethanes, and acrylics) and inorganic coatings (such as Polysiloxanes) is the most common method of mitigating corrosion of carbon steel. Incorporating the benefits of organic and inorganic hybrid (OIH) compounds for the designing of hybrid protective coatings is still challenging for industrial applications. There are advantages of inorganic coatings have, but purely inorganic siloxane-based coatings are difficult to use on industrial applications unless they are used at extremely low thicknesses (< 1-2 microns). Hence, most industrial applications try to have a combination of Polysiloxanes with organic compounds.  A hybrid coating possesses an organic section, which transports flexibility and impact resistance, and an inorganic section, which usually helps in the decreasing of porosity and increasing thermal stability and hardness. A number of polymers including polyethylene glycol and polyvinyl pyrrolidone have been reported to inhibit the corrosion mild steel in acidic media. However, reports on the effect of polyethylene oxide (PEO) or its blends on corrosion inhibition of metals is very scarce. Different composition of OIH coatings was synthesized by using silica sol-gel, epoxy, and PEO. The effect of different coating types on the corrosion behavior of carbon steel in harsh solution has been studied by weight loss and electrochemical measurements using Gamry 1000 Interface Potentiostat. Coating structures were investigated by SEM. İt revealed a considerable reduction in corrosion rate for coated sample. Based on these results, OIH coating prepared by epoxy-silica sol gel-PEO and epoxy-silica sol-gel exhibit had a %99.5 and %98 reduction of (Corrosion rate) CR compares to baseline. Cathodic Tafel constant (βc) shows that coatings change both Tafel constants but had more effect on the cathodic process. The evolution of the Potentiostatic scan with time displays stability in potential, some of them in a high value while the other in a low value which can be attributed to the formation of an oxide film covering substrate surface. The coated samples with the group of epoxy coating have a lower potential along with the time test, while the silica group shows higher in potential with respect to time.

Keywords: electrostatic, hybrid coating, corrosion tests, silica sol gel

Procedia PDF Downloads 92
145 Optimization of Culture Conditions of Paecilomyces Tenuipes, Entomopathogenic Fungi Inoculated into the Silkworm Larva, Bombyx Mori

Authors: Sung-Hee Nam, Kwang-Gill Lee, You-Young Jo, HaeYong Kweon

Abstract:

Entomopathogenic fungi is a Cordyceps species that is isolated from dead silkworm and cicada. Fungi on cicadas were described in old Chinese medicinal books and From ancient times, vegetable wasps and plant worms were widely known to have active substance and have been studied for pharmacological use. Among many fungi belonging to the genus Cordyceps, Cordyceps sinensis have been demonstrated to yield natural products possessing various biological activities and many bioactive components. Generally, It is commonly used to replenish the kidney and soothe the lung, and for the treatment of fatigue. Due to their commercial and economic importance, the demand for Cordyceps has been rapidly increased. However, a supply of Cordyceps specimen could not meet the increasing demand because of their sole dependence on field collection and habitat destruction. Because it is difficult to obtain many insect hosts in nature and the edibility of host insect needs to be verified in a pharmacological aspect. Recently, this setback was overcome that P. tenuipes was able to be cultivated in a large scale using silkworm as host. Pharmacological effects of P. tenuipes cultured on silkworm such as strengthening immune function, anti-fatigue, anti-tumor activity and controlling liver etc have been proved. They are widely commercialized. In this study, we attempted to establish a method for stable growth inhibition of P. tenuipes on silkworm hosts and an optimal condition for synnemata formation. To determine optimum culturing conditions, temperature and light conditions were varied. The length and number of synnemata was highest at 25℃ temperature and 100~300 lux illumination. On an average, the synnemata of wild P. tenuipes measures 70 ㎜ in length and 20 in number; those of the cultured strain were relatively shorter and more in number. The number of synnemata may have increased as a result of inoculating the host with highly concentrated conidia, while the length may have decreased due to limited nutrition per individual. It is not able that changes in light illumination cause morphological variations in the synnemata. However, regulation of only light and temperature could not produce stromata like perithecia, asci, and ascospores. Yamanaka reported that although a complete fruiting body can be produced under optimal culture conditions, it should be regarded as synnemata because it does not develop into an ascoma bearing ascospores.

Keywords: paecilomyces tenuipes, entomopathogenic fungi, silkworm larva, bombyx mori

Procedia PDF Downloads 299
144 The Effect of Framework Structure on N2O Formation over Cu-Based Zeolites during NH3-SCR Reactions

Authors: Ghodsieh Isapour Toutizad, Aiyong Wang, Joonsoo Han, Derek Creaser, Louise Olsson, Magnus Skoglundh, Hanna HaRelind

Abstract:

Nitrous oxide (N2O), which is generally formed as a byproduct of industrial chemical processes and fossil fuel combustion, has attracted considerable attention due to its destructive role in global warming and ozone layer depletion. From various developed technologies used for lean NOx reduction, the selective catalytic reduction (SCR) of NOx with ammonia is presently the most applied method. Therefore, the development of catalysts for efficient lean NOx reduction without forming N2O in the process, or only forming it to a very small extent from the exhaust gases is of crucial significance. One type of catalysts that nowadays are used for this aim are zeolite-based catalysts. It is owing to their remarkable catalytic performance under practical reaction conditions such as high thermal stability and high N2 selectivity. Among all zeolites, copper ion-exchanged zeolites, with CHA, MFI, and BEA framework structure (like SSZ-13, ZSM-5 and Beta, respectively), represent higher hydrothermal stability, high activity and N2 selectivity. This work aims at investigating the effect of the zeolite framework structure on the formation of N2O during NH3-SCR reaction conditions over three Cu-based zeolites ranging from small-pore to large-pore framework structure. In the zeolite framework, Cu exists in two cationic forms, that can catalyze the SCR reaction by activating NO to form NO+ and/or surface nitrate species. The nitrate species can thereafter react with NH3 to form another intermediate, ammonium nitrate, which seems to be one source for N2O formation at low temperatures. The results from in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS) indicate that during the NO oxidation step, mainly NO+ and nitrate species are formed on the surface of the catalysts. The intensity of the absorption peak attributed to NO+ species is higher for the Cu-CHA sample compared to the other two samples, indicating a higher stability of this species in small cages. Furthermore, upon the addition of NH3, through the standard SCR reaction conditions, absorption peaks assigned to N-H stretching and bending vibrations are building up. At the same time, negative peaks are evolving in the O-H stretching region, indicating blocking/replacement of surface OH-groups by NH3 and NH4+. By removing NH3 and adding NO2 to the inlet gas composition, the peaks in the N-H stretching and bending vibration regions show a decreasing trend in intensity, with the decrease being more pronounced for increasing pore size. It can probably be owing to the higher accumulation of ammonia species in the small-pore size zeolite compared to the other two samples. Furthermore, it is worth noting that the ammonia surface species are strongly bonded to the CHA zeolite structure, which makes it more difficult to react with NO2. To conclude, the framework structure of the zeolite seems to play an important role in the formation and reactivity of surface species relevant for the SCR process. Here we intend to discuss the connection between the zeolite structure, the surface species, and the formation of N2O during ammonia-SCR.

Keywords: fast SCR, nitrous oxide, NOx, standard SCR, zeolites

Procedia PDF Downloads 207
143 The Antioxidant Gel Mask Supplies Of Bitter Melon's Extract ( Momordica charantia Linn.)

Authors: N. S. Risqina, G. Edijanti, P. S. Nurita, L. Endang, R. A. Siti, R. Tri

Abstract:

Skin is an important and vital organs and also as a mirror of health and life. Facial skin care is one of the main emphasis to get the beautiful, healthy, and fresh skin. Potentially antioxidant phenolic compounds shows, antimutagen, antitumor, anti-inflammatory, and anti-cancer. Flavonoids are a group of polyphenolic compounds that have the nature of free radicals, inhibiting the oxidative and hydrolytic enzymes as well as anti-inflammatory. Bitter melon (Momordica charantia Linn) is a plant that contains flavonoids, and phenolic antioxidant activity. Bitter melon has strong antioxidant activity that can counteract the free radicals.These compounds can prevent free radicals that cause premature aging. Gel masks including depth cleansing is the cosmetics which work in depth and could raise the dead skin cells. Measurement of antioxidant activity of the extract and gel mask is done by using the immersion method of DPPH. IC50 value of ethanol extract of bitter melon fruit of 287.932 ppm. The preparation of gel mask bitter melon fruit extract, necessary to test the effectiveness of antioxidants using DPPH method is done by measuring the inhibition of DPPH and using UV spectrophotometer at the wavelength of maximum DPPH solution. Tests conducted at the beginning and end of the evaluation (day 0 and day 28). The purpose of this study is to determine the antioxidant activity of the bitter melon's extract and to determine the antioxidant activity of ethanol extract gel mask pare in varying concentrations, ie 1xIC100 (0.295%), 2xIC100 (0.590%) and 4xIC100 (1.180%). Evaluation of physical properties of the preparation on (Day-0,7,14,21, and 28) and evaluation of antioxidant activity (day 0 and 28). Data were analyzed using One Way ANOVA to determine differences in the physical properties of each formula. The statistical results showed that differences in the formula and storage time affects the adhesion, dispersive power, dry time and pH it is shown on a significant value of p <0.05, but longer storage does not affect the pH because the significance value p> 0,05. The antioxidant test showed that there are differences in antioxidant activity in all formulas. Measurement of antioxidant activity of bitter melon fruit extract gel mask on day 0 with a concentration of 0.295%, 0.590%, and 1.180%, respectively, are 124,209.277 ppm, ppm 83819.223 and 47323.592 ppm, whereas day 28 consecutive 130 411, 495 ppm, and 53239.806 95561.645 ppm ppm. The Conclusions drawn that there are antioxidant activity in preparation gel mask of bitter melon fruit extract. The antioxidant activity of bitter melon fruit extract gel mask on the day 0 with a concentration of 0.295%, 0.590%, and 1.180%, respectively, are 124,209.277 ppm, ppm 83819.223 and 47323.592 ppm, whereas on day 28 of antioxidant activity gel mask bitter melon fruit extract with a concentration of 0.295%, 0.590%, and 1.180% in succession, namely: 130,411.495 ppm, ppm 95561.645 and 53239.806 ppm.

Keywords: antioxdant, bitter melon, gel mask, IC50

Procedia PDF Downloads 449
142 A Computational Model of the Thermal Grill Illusion: Simulating the Perceived Pain Using Neuronal Activity in Pain-Sensitive Nerve Fibers

Authors: Subhankar Karmakar, Madhan Kumar Vasudevan, Manivannan Muniyandi

Abstract:

Thermal Grill Illusion (TGI) elicits a strong and often painful sensation of burn when interlacing warm and cold stimuli that are individually non-painful, excites thermoreceptors beneath the skin. Among several theories of TGI, the “disinhibition” theory is the most widely accepted in the literature. According to this theory, TGI is the result of the disinhibition or unmasking of the pain-sensitive HPC (Heat-Pinch-Cold) nerve fibers due to the inhibition of cold-sensitive nerve fibers that are responsible for masking HPC nerve fibers. Although researchers focused on understanding TGI throughexperiments and models, none of them investigated the prediction of TGI pain intensity through a computational model. Furthermore, the comparison of psychophysically perceived TGI intensity with neurophysiological models has not yet been studied. The prediction of pain intensity through a computational model of TGI can help inoptimizing thermal displays and understanding pathological conditions related to temperature perception. The current studyfocuses on developing a computational model to predict the intensity of TGI pain and experimentally observe the perceived TGI pain. The computational model is developed based on the disinhibition theory and by utilizing the existing popular models of warm and cold receptors in the skin. The model aims to predict the neuronal activity of the HPC nerve fibers. With a temperature-controlled thermal grill setup, fifteen participants (ten males and five females) were presented with five temperature differences between warm and cold grills (each repeated three times). All the participants rated the perceived TGI pain sensation on a scale of one to ten. For the range of temperature differences, the experimentally observed perceived intensity of TGI is compared with the neuronal activity of pain-sensitive HPC nerve fibers. The simulation results show a monotonically increasing relationship between the temperature differences and the neuronal activity of the HPC nerve fibers. Moreover, a similar monotonically increasing relationship is experimentally observed between temperature differences and the perceived TGI intensity. This shows the potential comparison of TGI pain intensity observed through the experimental study with the neuronal activity predicted through the model. The proposed model intends to bridge the theoretical understanding of the TGI and the experimental results obtained through psychophysics. Further studies in pain perception are needed to develop a more accurate version of the current model.

Keywords: thermal grill Illusion, computational modelling, simulation, psychophysics, haptics

Procedia PDF Downloads 140
141 Effects of Starvation, Glucose Treatment and Metformin on Resistance in Chronic Myeloid Leukemia Cells

Authors: Nehir Nebioglu

Abstract:

Chemotherapy is widely used for the treatment of cancer. Doxorubicin is an anti-cancer chemotherapy drug that is classified as an anthracycline antibiotic. Antitumor antibiotics consist of natural products produced by species of the soil fungus Streptomyces. These drugs act in multiple phases of the cell cycle and are known cell-cycle specific. Although DOX is a precious clinical antineoplastic agent, resistance is also a problem that limits its utility besides cardiotoxicity problem. The drug resistance of cancer cells results from multiple factors including individual variation, genetic heterogeneity within a tumor, and cellular evolution. The mechanism of resistance is thought to involve, in particular, ABCB1 (MDR1, Pgp) and ABCC1 (MRP1) as well as other transporters. Several studies on DOX-resistant cell lines have shown that resistance can be overcome by an inhibition of ABCB1, ABCC1, and ABCC2. This study attempts to understand the effects of different concentration levels of glucose treatment and starvation on the proliferation of Doxorubicin resistant cancer cells lines. To understand the effect of starvation, K562/Dox and K562 cell lines were treated with 0, 5 nM, 50 nM, 500 nM, 5 uM and 50 uM Dox concentrations in both starvation and normal medium conditions. In addition to this, to interpret the effect of glucose treatment, different concentrations (0, 1 mM, 5 mM, 25 mM) of glucose were applied to Dox-treated (with 0, 5 nM, 50 nM, 500 nM, 5 uM and 50 uM) K562/Dox and K652 cell lines. All results show significant decreasing in the cell count of K562/Dox, when cells were starved. However, while proliferation of K562/Dox lines decrease is associated with the increasingly applied Dox concentration, K562/Dox starved ones remain at the same proliferation level. Thus, the results imply that an amount of K562/Dox lines gain starvation resistance and remain resistant. Furthermore, for K562/Dox, there is no clear effect of glucose treatment in terms of cell proliferation. In the presence of a moderate level of glucose (5 mM), proliferation increases compared to other concentration of glucose for each different Dox application. On the other hand, a significant increase in cell proliferation in moderate level of glucose is only observed in 5 uM Dox concentration. The moderate concentration level of Dox can be examined in further studies. For the high amount of glucose (25 mM), cell proliferation levels are lower than moderate glucose application. The reason could be high amount of glucose may not be absorbable by cells. Also, in the presence of low amount of glucose, proliferation is decreasing in an orderly manner of increase in Dox concentration. This situation can be explained by the glucose depletion -Warburg effect- in the literature.

Keywords: drug resistance, cancer cells, chemotherapy, doxorubicin

Procedia PDF Downloads 153
140 Anti-Neuroinflammatory and Anti-Apoptotic Efficacy of Equol, against Lipopolysaccharide Activated Microglia and Its Neurotoxicity

Authors: Lalita Subedi, Jae Kyoung Chae, Yong Un Park, Cho Kyo Hee, Lee Jae Hyuk, Kang Min Cheol, Sun Yeou Kim

Abstract:

Neuroinflammation may mediate the relationship between low levels of estrogens and neurodegenerative disease. Estrogens are neuroprotective and anti-inflammatory in neurodegenerative disease models. Due to the long term side effects of estrogens, researches have been focused on finding an effective phytoestrogens for biological activities. Daidzein present in soybeans and its active metabolite equol (7-hydroxy-3-(4'-hydroxyphenyl)-chroman) bears strong antioxidant and anticancer showed more potent anti-inflammatory and neuroprotective role in neuroinflammatory model confirmed its in vitro activity with molecular mechanism through NF-κB pathway. Three major CNS cells Microglia (BV-2), Astrocyte (C6), Neuron (N2a) were used to find the effect of equol in inducible nitric oxide synthase (iNOS), cyclooxygenase (COX-2), MAPKs signaling proteins, apoptosis related proteins by western blot analysis. Nitric oxide (NO) and prostaglandin E2 (PGE2) was measured by the Gries method and ELISA, respectively. Cytokines like tumor necrosis factor-α (TNF-α) and IL-6 were also measured in the conditioned medium of LPS activated cells with or without equol. Equol inhibited the NO production, PGE-2 production and expression of COX-2 and iNOS in LPS-stimulated microglial cells at a dose dependent without any cellular toxicity. At the same time Equol also showed promising effect in modulation of MAPK’s and nuclear factor kappa B (NF-κB) expression with significant inhibition of the production of proinflammatory cytokine like interleukin -6 (IL-6), and tumor necrosis factor -α (TNF-α). Additionally, it inhibited the LPS activated microglia-induced neuronal cell death by downregulating the apoptotic phenomenon in neuronal cells. Furthermore, equol increases the production of neurotrophins like NGF and increase the neurite outgrowth as well. In conclusion the natural daidzein metabolite equol are more active than daidzein, which showed a promising effectiveness as an anti-neuroinflammatory and neuroprotective agent via downregulating the LPS stimulated microglial activation and neuronal apoptosis. This work was supported by Brain Korea 21 Plus project and High Value-added Food Technology Development Program 114006-4, Ministry of Agriculture, Food and Rural Affairs.

Keywords: apoptosis, equol, neuroinflammation, phytoestrogen

Procedia PDF Downloads 343
139 In Vivo Antiulcer and Anti-Helicobacter pylori Activity of Geraniol on Acetic Acid plus Helicobacter pylori Induced Ulcer in Rats

Authors: Subrat Kumar Bhattamisra, Vivian Lee Yean Yan, Chin Koh Lee, Chew Hui Kuean, Yun Khoon Liew, Mayuren Candasamy

Abstract:

Geraniol, an acyclic monoterpenoid is the main active constituent in the essential oils of rose and palmorosa. Antioxidant, antibacterial, anticancer and antiulcer activity of geraniol was reported by many researchers. The present investigation was designed to study in vivo antiulcer and anti-Helicobacter pylori activity of geraniol. Antiulcer and anti-H. pylori activity of geraniol was evaluated on acetic acid plus H. pylori induced ulcer in rats. Acetic acid (0.03 mL) was injected to the sub-serosal layer of the stomach through laparotomy under anaesthesia. Orogastric inoculation of H. pylori (ATCC 43504) was done twice daily for 7 days. Geraniol (15 and 30 mg/kg), vehicle and standard drugs (Amoxicillin, 50 mg/kg; clarithromycin, 25 mg/kg & omeprazole, 20 mg/kg) was administered twice daily for 14 days. Antiulcer activity of geraniol was examined by the determination of gastric ulcer index, measuring the volume of gastric juice, pH and total acidity, myeloperoxidase activity and histopathological examination. Histopathological investigation for the presence of inflammation, white blood cell infiltration, edema, the mucosal damage was studied. The presence of H. pylori was detected by placing a biopsy sample from antral part of the stomach into rapid urease test. Ulcer index in H. pylori inoculated control group was 4.13 ± 0.85 and was significantly (P < 0.05) lowered in geraniol (30 mg/kg) and reference drug treated group. Geraniol increase the pH of the gastric juice (2.18 ± 0.13 in control vs. 4.14 ± 0.57 in geraniol 30mg/kg) and lower total acidity significantly (P < 0.01) in geraniol (15 & 30 mg/kg). Myeloperoxidase (MPO) activity was measured in stomach homogenate of all the groups. H. pylori control group has significant (P < 0.05) increase in MPO activity compared to normal control group. Geraniol (30 mg/kg) was showed significant (P < 0.05) and most effective among all the groups. Histopathological examination of rat stomach was scored and the total score for H. pylori control group was 8. After geraniol (30 mg/kg) and reference drug treatment, the histopathological score was significantly decreased and it was observed to be 3.5 and 2.0 respectively. Percentage inhibition of H. pylori infection in geraniol (30 mg/kg) and reference drug were found to be 40% and 50% respectively whereas, 100% infection in H. pylori control group was observed. Geraniol exhibited significant antiulcer and anti- H. pylori activity in the rats. Thus, geraniol has the potential for the further development as an effective medication in treating H. pylori associated ulcer.

Keywords: geraniol, helicobacter pylori atcc 43504, myeloperoxidase, ulcer

Procedia PDF Downloads 318
138 Refractory T-Cell Prolymphocytic Leukemia with JAK3 Mutation: In Vitro and Clinical Synergy of Tofacitinib and Ruxolitinib

Authors: Mike Wei, Nebu Koshy, Koen van Besien, Giorgio Inghirami, Steven M. Horwitz

Abstract:

T-cell prolymphocytic leukemia (T-PLL) is a rare hematologic disease characterized by a T-cell phenotype, rapid progression, and poor prognosis with median survival of less than a year. Alemtuzumab-based chemotherapy has increased the rate of complete remissions but these are often short-lived, and allogeneic transplant is considered the only curative therapy. In recent studies, JAK3 activating mutations have been identified in T-cell cancers, with T-PLL having the highest rate of JAK3 mutations (30 – 42%). As such, T-PLL is a model disease for evaluating the utility of JAK3 inhibitors. We present a case of a 64-year-old man with relapsed-refractory T-PLL. He was initially treated with alemtuzumab and obtained complete response and was consolidated with matched unrelated donor stem cell transplant. His disease stayed in remission for approximately 1.5 years before relapse, which was then treated with a clinical trial of romidepsin-lenalidomide (partial responses then progression at 6 months) and later alemtuzumab. Due to complications of myelosuppression and CMV reactivation, his treatment was interrupted leading to disease progression. The doubling time of lymphocyte count was approximately 20 days and over a span of 60 days the lymphocyte count rose from 8 x 109/L to 68 x 109/L. Exon sequencing showed a JAK3 mutation. The patient consented to and was treated with FDA-approved tofacitinib (initially 5 mg BID, increased to 10 mg BID after 15 days of treatment). An initial decrease in lymphocyte count was followed by progression. In vitro treatment of the patient’s cells showed modest effects of tofacitinib and ruxolitinib as single agents, in the range of doxorubicin, but synergy between the agents. After 40 days of treatment with tofacitinib and with a lymphocyte count of 150 x 109/L, ruxolitinib (5mg BID) was added. Over the 60 days since dual inhibition was started, the lymphocyte count has stabilized. The patient has remained completely asymptomatic during treatment with tofacitinib and ruxolitinib. Neutrophil count has remained normal. Platelet count and hemoglobin have however declined from ~50 x109/L to ~30 x109/L and from 11 g/dL to 8.1 g/dL respectively, since the introduction of ruxolitinib. The stabilization in lymphocyte count confirms the clinical activity of JAK inhibitors in T-PLL as suggested by the presence of JAK3 mutations and by in-vitro assays. It also suggests clinical synergy between ruxolitinib and tofacitinib in this setting. Prospective studies of JAK inhibitors in PLL patients with formal dose-finding studies are needed.

Keywords: tofacitinib, ruxolitinib, T-cell prolymphocytic leukemia, JAK3

Procedia PDF Downloads 292
137 Sustainable Nanoengineering of Copper Oxide: Harnessing Its Antimicrobial and Anticancer Capabilities

Authors: Yemane Tadesse Gebreslassie, Fisseha Guesh Gebremeskel

Abstract:

Nanotechnology has made remarkable advancements in recent years, revolutionizing various scientific fields, industries, and research institutions through the utilization of metal and metal oxide nanoparticles. Among these nanoparticles, copper oxide nanoparticles (CuO NPs) have garnered significant attention due to their versatile properties and wide-range applications, particularly, as effective antimicrobial and anticancer agents. CuO NPs can be synthesized using different methods, including physical, chemical, and biological approaches. However, conventional chemical and physical approaches are expensive, resource-intensive, and involve the use of hazardous chemicals, which can pose risks to human health and the environment. In contrast, biological synthesis provides a sustainable and cost-effective alternative by eliminating chemical pollutants and allowing for the production of CuO NPs of tailored sizes and shapes. This comprehensive review focused on the green synthesis of CuO NPs using various biological resources, such as plants, microorganisms, and other biological derivatives. Current knowledge and recent trends in green synthesis methods for CuO NPs are discussed, with a specific emphasis on their biomedical applications, particularly in combating cancer and microbial infections. This review highlights the significant potential of CuO NPs in addressing these diseases. By capitalizing on the advantages of biological synthesis, such as environmental safety and the ability to customize nanoparticle characteristics, CuO NPs have emerged as promising therapeutic agents for a wide range of conditions. This review presents compelling findings, demonstrating the remarkable achievements of biologically synthesized CuO NPs as therapeutic agents. Their unique properties and mechanisms enable effective combating against cancer cells and various harmful microbial infections. CuO NPs exhibit potent anticancer activity through diverse mechanisms, including induction of apoptosis, inhibition of angiogenesis, and modulation of signaling pathways. Additionally, their antimicrobial activity manifests through various mechanisms, such as disrupting microbial membranes, generating reactive oxygen species, and interfering with microbial enzymes. This review offers valuable insights into the substantial potential of biologically synthesized CuO NPs as an alternative approach for future therapeutic interventions against cancer and microbial infections.

Keywords: copper oxide nanoparticles, green synthesis, nanotechnology, microbial infection

Procedia PDF Downloads 31
136 Identification of a Lead Compound for Selective Inhibition of Nav1.7 to Treat Chronic Pain

Authors: Sharat Chandra, Zilong Wang, Ru-Rong Ji, Andrey Bortsov

Abstract:

Chronic pain (CP) therapeutic approaches have limited efficacy. As a result, doctors are prescribing opioids for chronic pain, leading to opioid overuse, abuse, and addiction epidemic. Therefore, the development of effective and safe CP drugs remains an unmet medical need. Voltage-gated sodium (Nav) channels act as cardiovascular and neurological disorder’s molecular targets. Nav channels selective inhibitors are hard to design because there are nine closely-related isoforms (Nav1.1-1.9) that share the protein sequence segments. We are targeting the Nav1.7 found in the peripheral nervous system and engaged in the perception of pain. The objective of this project was to screen a 1.5 million compound library for identification of inhibitors for Nav1.7 with analgesic effect. In this study, we designed a protocol for identification of isoform-selective inhibitors of Nav1.7, by utilizing the prior information on isoform-selective antagonists. First, a similarity search was performed; then the identified hits were docked into a binding site on the fourth voltage-sensor domain (VSD4) of Nav1.7. We used the FTrees tool for similarity searching and library generation; the generated library was docked in the VSD4 domain binding site using FlexX and compounds were shortlisted using a FlexX score and SeeSAR hyde scoring. Finally, the top 25 compounds were tested with molecular dynamics simulation (MDS). We reduced our list to 9 compounds based on the MDS root mean square deviation plot and obtained them from a vendor for in vitro and in vivo validation. Whole-cell patch-clamp recordings in HEK-293 cells and dorsal root ganglion neurons were conducted. We used patch pipettes to record transient Na⁺ currents. One of the compounds reduced the peak sodium currents in Nav1.7-HEK-293 stable cell line in a dose-dependent manner, with IC50 values at 0.74 µM. In summary, our computer-aided analgesic discovery approach allowed us to develop pre-clinical analgesic candidate with significant reduction of time and cost.

Keywords: chronic pain, voltage-gated sodium channel, isoform-selective antagonist, similarity search, virtual screening, analgesics development

Procedia PDF Downloads 98
135 Antibacterial Effect of Silver Diamine Fluoride Incorporated in Fissure Sealants

Authors: Nélio Veiga, Paula Ferreira, Tiago Correia, Maria J. Correia, Carlos Pereira, Odete Amaral, Ilídio J. Correia

Abstract:

Introduction: The application of fissure sealants is considered to be an important primary prevention method used in dental medicine. However, the formation of microleakage gaps between tooth enamel and the fissure sealant applied is one of the most common reasons of dental caries development in teeth with fissure sealants. The association between various dental biomaterials may limit the major disadvantages and limitations of biomaterials functioning in a complementary manner. The present study consists in the incorporation of a cariostatic agent – silver diamine fluoride (SDF) – in a resin-based fissure sealant followed by the study of release kinetics by spectrophotometry analysis of the association between both biomaterials and assessment of the inhibitory effect on the growth of the reference bacterial strain Streptococcus mutans (S. mutans) in an in vitro study. Materials and Methods: An experimental in vitro study was designed consisting in the entrapment of SDF (Cariestop® 12% and 30%) into a commercially available fissure sealant (Fissurit®), by photopolymerization and photocrosslinking. The same sealant, without SDF was used as a negative control. The effect of the sealants on the growth of S. mutans was determined by the presence of bacterial inhibitory halos in the cultures at the end of the incubation period. In order to confirm the absence of bacteria in the surface of the materials, Scanning Electron Microscopy (SEM) characterization was performed. Also, to analyze the release profile of SDF along time, spectrophotometry technique was applied. Results: The obtained results indicate that the association of SDF to a resin-based fissure sealant may be able to increase the inhibition of S. mutans growth. However, no SDF release was noticed during the in vitro release studies and no statistical significant difference was verified when comparing the inhibitory halo sizes obtained for test and control group.  Conclusions: In this study, the entrapment of SDF in the resin-based fissure sealant did not potentiate the antibacterial effect of the fissure sealant or avoid the immediate development of dental caries. The development of more laboratorial research and, afterwards, long-term clinical data are necessary in order to verify if this association between these biomaterials is effective and can be considered for being used in oral health management. Also, other methodologies for associating cariostatic agents and sealant should be addressed.

Keywords: biomaterial, fissure sealant, primary prevention, silver diamine fluoride

Procedia PDF Downloads 235
134 Green and Cost-Effective Biofabrication of Copper Oxide Nanoparticles: Exploring Antimicrobial and Anticancer Applications

Authors: Yemane Tadesse Gebreslassie, Fisseha Guesh Gebremeskel

Abstract:

Nanotechnology has made remarkable advancements in recent years, revolutionizing various scientific fields, industries, and research institutions through the utilization of metal and metal oxide nanoparticles. Among these nanoparticles, copper oxide nanoparticles (CuO NPs) have garnered significant attention due to their versatile properties and wide-range applications, particularly, as effective antimicrobial and anticancer agents. CuO NPs can be synthesized using different methods, including physical, chemical, and biological approaches. However, conventional chemical and physical approaches are expensive, resource-intensive, and involve the use of hazardous chemicals, which can pose risks to human health and the environment. In contrast, biological synthesis provides a sustainable and cost-effective alternative by eliminating chemical pollutants and allowing for the production of CuO NPs of tailored sizes and shapes. This comprehensive review focused on the green synthesis of CuO NPs using various biological resources, such as plants, microorganisms, and other biological derivatives. Current knowledge and recent trends in green synthesis methods for CuO NPs are discussed, with a specific emphasis on their biomedical applications, particularly in combating cancer and microbial infections. This review highlights the significant potential of CuO NPs in addressing these diseases. By capitalizing on the advantages of biological synthesis, such as environmental safety and the ability to customize nanoparticle characteristics, CuO NPs have emerged as promising therapeutic agents for a wide range of conditions. This review presents compelling findings, demonstrating the remarkable achievements of biologically synthesized CuO NPs as therapeutic agents. Their unique properties and mechanisms enable effective combating against cancer cells and various harmful microbial infections. CuO NPs exhibit potent anticancer activity through diverse mechanisms, including induction of apoptosis, inhibition of angiogenesis, and modulation of signaling pathways. Additionally, their antimicrobial activity manifests through various mechanisms, such as disrupting microbial membranes, generating reactive oxygen species, and interfering with microbial enzymes. This review offers valuable insights into the substantial potential of biologically synthesized CuO NPs as an alternative approach for future therapeutic interventions against cancer and microbial infections.

Keywords: biological synthesis, copper oxide nanoparticles, microbial infection, nanotechnology

Procedia PDF Downloads 32
133 Whey Protein: A Noval Protective Agent against Oto-Toxicity Induced by Cis-Platin in Male Rat

Authors: Eitedal Daoud, Reda M.Daoud, Khaled Abdel-Wahhab, Maha M.Saber, Lobna Saber

Abstract:

Background: Cis-platin is a widely used chemotherapeutic drug to treat many malignant disorders including head and neck malignancies. Oto-nephrotxicity is an important and dose - limiting side effect of cis - platin therapy. Nowadays, more attention had been paid to oto-toxicity caused with cis-platin. Aim of the Work: This study was designed to investigate the potential protective effect of Whey protein (WP) against cis-platin induced ototoxicity compared to the effect of N-acetylcysteine (NAC) in rats. Methodology: Male albino rats were randomly divided into 6 groups: untreated rats (control), rats orally treated with whey protein (1g/kg b.w/day) for seven executive days, rats treated orally with N-acetylcysteine (500 mg/kgb.w /day) for seven executive days, rates intoxicated intraperitoneal (ip) with cis- platin (10 mg/kgb.w. once), rats treated with whey protein (1g/kgb.w./day) for seven executive days) followed by one injection (ip) of cis-platin(10 mg/kg b.w.) one hour after the last oral administration of whey protein, rats treated with N- acetylcysteine (for seven executive days followed by one injection (ip) of cis-platin (10 mg/kgb.w) one hour after the last oral administration of N-acetylcysteine. The organ of Corti, the stria vascularis and spiral ganglia were visualized by light microscopy at different magnifications. Results: Cis-platin intoxicated animals showed a significant decrease in serum level of total antioxidant capacity (TAC),with inhibition in the activity of serum glutathione-s transferase(GST) and paraoxonnase-1 (PON-1) in comparison with control. Group treated with either NAC or WP with cis-platin showed significant elevation in the activity of both GST & PON-1 with increased serum level of TAC when compared with cis-platin intoxicated rats. Animals treated with NAC or WP with cis-platin compared to those treated with cis-platin alone showed marked degree of improvement towards control rats as there was significant drop in the serum level of cortecosterone, nitric oxide (NO), and melandialdehyde (MDA).Histopathologic, in NAC pretreated group there was no changes in stria vascularis or spiral ganglia. In group pretreated with WP, there was no histopathologic alteration detected in the organ of Corti and Reissers membrane but oedema and haemorrhage were founded in the stria vascularis in small focal manner. Conclusion: Our finding showed that Whey protein is a natural dietary supplement product proved its ability of protection of anti-oxidant system and the cochlea against cis-platin induced ototoxicity.

Keywords: anti-oxidant, cis-platin, N-acetylcysteine, ototoxicity, whey protein

Procedia PDF Downloads 499
132 In silico Designing of Imidazo [4,5-b] Pyridine as a Probable Lead for Potent Decaprenyl Phosphoryl-β-D-Ribose 2′-Epimerase (DprE1) Inhibitors as Antitubercular Agents

Authors: Jineetkumar Gawad, Chandrakant Bonde

Abstract:

Tuberculosis (TB) is a major worldwide concern whose control has been exacerbated by HIV, the rise of multidrug-resistance (MDR-TB) and extensively drug resistance (XDR-TB) strains of Mycobacterium tuberculosis. The interest for newer and faster acting antitubercular drugs are more remarkable than any time. To search potent compounds is need and challenge for researchers. Here, we tried to design lead for inhibition of Decaprenyl phosphoryl-β-D-ribose 2′-epimerase (DprE1) enzyme. Arabinose is an essential constituent of mycobacterial cell wall. DprE1 is a flavoenzyme that converts decaprenylphosphoryl-D-ribose into decaprenylphosphoryl-2-keto-ribose, which is intermediate in biosynthetic pathway of arabinose. Latter, DprE2 converts keto-ribose into decaprenylphosphoryl-D-arabinose. We had a selection of 23 compounds from azaindole series for computational study, and they were drawn using marvisketch. Ligands were prepared using Maestro molecular modeling interface, Schrodinger, v10.5. Common pharmacophore hypotheses were developed by applying dataset thresholds to yield active and inactive set of compounds. There were 326 hypotheses were developed. On the basis of survival score, ADRRR (Survival Score: 5.453) was selected. Selected pharmacophore hypotheses were subjected to virtual screening results into 1000 hits. Hits were prepared and docked with protein 4KW5 (oxydoreductase inhibitor) was downloaded in .pdb format from RCSB Protein Data Bank. Protein was prepared using protein preparation wizard. Protein was preprocessed, the workspace was analyzed using force field OPLS 2005. Glide grid was generated by picking single atom in molecule. Prepared ligands were docked with prepared protein 4KW5 using Glide docking. After docking, on the basis of glide score top-five compounds were selected, (5223, 5812, 0661, 0662, and 2945) and the glide docking score (-8.928, -8.534, -8.412, -8.411, -8.351) respectively. There were interactions of ligand and protein, specifically HIS 132, LYS 418, TRY 230, ASN 385. Pi-pi stacking was observed in few compounds with basic Imidazo [4,5-b] pyridine ring. We had basic azaindole ring in parent compounds, but after glide docking, we received compounds with Imidazo [4,5-b] pyridine as a basic ring. That might be the new lead in the process of drug discovery.

Keywords: DprE1 inhibitors, in silico drug designing, imidazo [4, 5-b] pyridine, lead, tuberculosis

Procedia PDF Downloads 129
131 Analysis of Taxonomic Compositions, Metabolic Pathways and Antibiotic Resistance Genes in Fish Gut Microbiome by Shotgun Metagenomics

Authors: Anuj Tyagi, Balwinder Singh, Naveen Kumar B. T., Niraj K. Singh

Abstract:

Characterization of diverse microbial communities in specific environment plays a crucial role in the better understanding of their functional relationship with the ecosystem. It is now well established that gut microbiome of fish is not the simple replication of microbiota of surrounding local habitat, and extensive species, dietary, physiological and metabolic variations in fishes may have a significant impact on its composition. Moreover, overuse of antibiotics in human, veterinary and aquaculture medicine has led to rapid emergence and propagation of antibiotic resistance genes (ARGs) in the aquatic environment. Microbial communities harboring specific ARGs not only get a preferential edge during selective antibiotic exposure but also possess the significant risk of ARGs transfer to other non-resistance bacteria within the confined environments. This phenomenon may lead to the emergence of habitat-specific microbial resistomes and subsequent emergence of virulent antibiotic-resistant pathogens with severe fish and consumer health consequences. In this study, gut microbiota of freshwater carp (Labeo rohita) was investigated by shotgun metagenomics to understand its taxonomic composition and functional capabilities. Metagenomic DNA, extracted from the fish gut, was subjected to sequencing on Illumina NextSeq to generate paired-end (PE) 2 x 150 bp sequencing reads. After the QC of raw sequencing data by Trimmomatic, taxonomic analysis by Kraken2 taxonomic sequence classification system revealed the presence of 36 phyla, 326 families and 985 genera in the fish gut microbiome. At phylum level, Proteobacteria accounted for more than three-fourths of total bacterial populations followed by Actinobacteria (14%) and Cyanobacteria (3%). Commonly used probiotic bacteria (Bacillus, Lactobacillus, Streptococcus, and Lactococcus) were found to be very less prevalent in fish gut. After sequencing data assembly by MEGAHIT v1.1.2 assembler and PROKKA automated analysis pipeline, pathway analysis revealed the presence of 1,608 Metacyc pathways in the fish gut microbiome. Biosynthesis pathways were found to be the most dominant (51%) followed by degradation (39%), energy-metabolism (4%) and fermentation (2%). Almost one-third (33%) of biosynthesis pathways were involved in the synthesis of secondary metabolites. Metabolic pathways for the biosynthesis of 35 antibiotic types were also present, and these accounted for 5% of overall metabolic pathways in the fish gut microbiome. Fifty-one different types of antibiotic resistance genes (ARGs) belonging to 15 antimicrobial resistance (AMR) gene families and conferring resistance against 24 antibiotic types were detected in fish gut. More than 90% ARGs in fish gut microbiome were against beta-lactams (penicillins, cephalosporins, penems, and monobactams). Resistance against tetracycline, macrolides, fluoroquinolones, and phenicols ranged from 0.7% to 1.3%. Some of the ARGs for multi-drug resistance were also found to be located on sequences of plasmid origin. The presence of pathogenic bacteria and ARGs on plasmid sequences suggested the potential risk due to horizontal gene transfer in the confined gut environment.

Keywords: antibiotic resistance, fish gut, metabolic pathways, microbial diversity

Procedia PDF Downloads 112
130 Suture Biomaterials Development from Natural Fibers: Muga Silk (Antheraea assama) and Ramie (Boehmeria nivea)

Authors: Raghuram Kandimalla, Sanjeeb Kalita, Bhaswati Choudhury, Jibon Kotoky

Abstract:

The quest for developing an ideal suture material prompted our interest to develop a novel suture with advantageous characteristics to market available ones. We developed novel suture biomaterial from muga silk (Antheraea assama) and ramie (Boehmeria nivea) plant fiber. Field emission scanning electron microscopy (FE-SEM), energy-dispersive X-ray spectroscopy (EDX), attenuated total reflection fourier transform infrared spectroscopy (ATR-FTIR) and thermo gravimetric analysis (TGA) results revealed the physicochemical properties of the fibers which supports the suitability of fibers for suture fabrication. Tensile properties of the prepared sutures were comparable with market available sutures and it found to be biocompatible towards human erythrocytes and nontoxic to mammalian cells. The prepared sutures completely healed the superficial deep wound incisions within seven days in adult male wister rats leaving no rash and scar. Histopathology studies supports the wound healing ability of sutures, as rapid synthesis of collagen, connective tissue and other skin adnexal structures were observed within seven days of surgery. Further muga suture surface modified by exposing the suture to oxygen plasma which resulted in formation of nanotopography on suture surface. Broad spectrum antibiotic amoxicillin was functionalized on the suture surface to prepare an advanced antimicrobial muga suture. Surface hydrophilicity induced by oxygen plasma results in an increase in drug-impregnation efficiency of modified muga suture by 16.7%. In vitro drug release profiles showed continuous and prolonged release of amoxicillin from suture up to 336 hours. The advanced muga suture proves to be effective against growth inhibition of Staphylococcus aureus and Escherichia coli, whereas normal muga suture offers no antibacterial activity against both types of bacteria. In vivo histopathology studies and colony-forming unit count data revealed accelerated wound healing activity of advanced suture over normal one through rapid synthesis and proliferation of collagen, hair follicle and connective tissues.

Keywords: sutures, biomaterials, silk, Ramie

Procedia PDF Downloads 290