Search results for: keratin protein fiber

Authors: Sanjay Shukla, A. Nayak, R. K. Sharma, A. P. Singh, S. P. Tiwari

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Excessive use of antibiotics is a major problem in the treatment of wounds and other chronic infections, and antibiotic treatment is frequently non-curative, thus alternative treatment is necessary. Phage therapy is considered one of the most promising approaches to treat multi-drug resistant bacterial pathogens. Infections caused by Staphylococcus aureus are very efficiently controlled with phage cocktails, containing a different individual phages lysate infecting a majority of known pathogenic S. aureus strains. The aim of the present study was to evaluate the efficacy of a purified phage cocktail for prophylactic as well as therapeutic application in mouse model and in large animals with chronic septic infection of wounds. A total of 150 sewage samples were collected from various livestock farms. These samples were subjected for the isolation of bacteriophage by the double agar layer method. A total of 27 sewage samples showed plaque formation by producing lytic activity against S. aureus in the double agar overlay method out of 150 sewage samples. In TEM, recovered isolates of bacteriophages showed hexagonal structure with tail fiber. In the bacteriophage (ØVS) had an icosahedral symmetry with the head size 52.20 nm in diameter and long tail of 109 nm. Head and tail were held together by connector and can be classified as a member of the Myoviridae family under the order of Caudovirale. Recovered bacteriophage had shown the antibacterial activity against the S. aureus in vitro. Cocktail (ØVS1, ØVS5, ØVS9, and ØVS 27) of phage lysate were tested to know in vivo antibacterial activity as well as the safety profile. Result of mice experiment indicated that the bacteriophage lysate were very safe, did not show any appearance of abscess formation, which indicates its safety in living system. The mice were also prophylactically protected against S. aureus when administered with cocktail of bacteriophage lysate just before the administration of S. aureuswhich indicates that they are good prophylactic agent. The S. aureusinoculated mice were completely recovered by bacteriophage administration with 100% recovery, which was very good as compere to conventional therapy. In the present study, ten chronic cases of the wound were treated with phage lysate, and follow up of these cases was done regularly up to ten days (at 0, 5, and 10 d). The result indicated that the six cases out of ten showed complete recovery of wounds within 10 d. The efficacy of bacteriophage therapy was found to be 60% which was very good as compared to the conventional antibiotic therapy in chronic septic wounds infections. Thus, the application of lytic phage in single dose proved to be innovative and effective therapy for the treatment of septic chronic wounds.

Keywords: phage therapy, S aureus, antimicrobial resistance, lytic phage, and bacteriophage

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Authors: Bui Phuong Linh, Yuki Sakakibara, Ryuto Tanaka, Elizabeth H. Pigney, Taishi Hashiguchi

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NASH is an increasingly prevalent chronic liver disease that can progress to hepatocellular carcinoma and now is attracting interest worldwide. The STAM™ model is a clinically-correlated murine NASH model which shows the same pathological progression as NASH patients and has been widely used for pharmacological and basic research. The multiple parallel hits hypothesis suggests abnormalities in adipocytokines, intestinal microflora, and endotoxins are intertwined and could contribute to the development of NASH. In fact, NASH patients often exhibit gut dysbiosis and dysfunction in adipose tissue and metabolism. However, the analysis of the STAM™ model has only focused on the liver. To clarify whether the STAM™ model can also mimic multiple pathways of NASH progression, we analyzed the organ crosstalk interactions between the liver and the gut and the phenotype of adipose tissue in the STAM™ model. NASH was induced in male mice by a single subcutaneous injection of 200 µg streptozotocin 2 days after birth and feeding with high-fat diet after 4 weeks of age. The mice were sacrificed at NASH stage. Colon samples were snap-frozen in liquid nitrogen and stored at -80˚C for tight junction-related protein analysis. Adipose tissue was prepared into paraffin blocks for HE staining. Blood adiponectin was analyzed to confirm changes in the adipocytokine profile. Tight junction-related proteins in the intestine showed that expression of ZO-1 decreased with the progression of the disease. Increased expression of endotoxin in the blood and decreased expression of Adiponectin were also observed. HE staining revealed hypertrophy of adipocytes. Decreased expression of ZO-1 in the intestine of STAM™ mice suggests the occurrence of leaky gut, and abnormalities in adipocytokine secretion were also observed. Together with the liver, phenotypes in these organs are highly similar to human NASH patients and might be involved in the pathogenesis of NASH.

Keywords: Non-alcoholic steatohepatitis, hepatocellular carcinoma, fibrosis, organ crosstalk, leaky gut

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Authors: Ayhan Varol, Aysenur Ugurlu

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Due to the depletion of fossil fuels and climate change, there is a rising interest in renewable energy sources. In this concept, a wide range of biomass (energy crops, animal manure, solid wastes, etc.) are used for energy production. There has been a growing interest in biomethane production from biomass. Biomethane production from organic wastes is a promising alternative for waste management by providing organic matter stabilization. Anaerobic digestion of organic material produces biogas, and organic substrate is degraded into a more stable material. Therefore, anaerobic digestion technology helps reduction of carbon emissions and produces renewable energy. The hydraulic retention time (HRT) and organic loading rate (OLR), as well as TS (VS) loadings, influences the anaerobic digestion of organic wastes significantly. The optimum range for HRT varies between 15 days to 30 days, whereas OLR differs between 0.5 to 5 g/L.d depending on the substrate type and its lipid, protein and carbohydrate contents. The organic wastes have biogas production potential through anaerobic digestion. In this study, biomethane production potential of wastes like sugar beet bagasse, agricultural residues, food wastes, olive mill pulp, and dairy manure having different characteristics was investigated in mesophilic CSTR reactor, and their performances were compared. The reactor was mixed in order to provide homogenized content at a rate of 80 rpm. The organic matter content of these wastes was between 85 to 94 % with 61% (olive pulp) to 22 % (food waste) dry matter content. The hydraulic retention time changed between 20-30 days. High biogas productions, 13.45 to 5.70 mL/day, were achieved from the wastes studied when operated at 9 to 10.5% TS loadings where OLR varied between 2.92 and 3.95 gVS/L.day. The results showed that food wastes have higher specific methane production rate and volumetric methane production potential than the other wastes studied, under the similar OLR values. The SBP was 680, 585, 540, 390 and 295 mL/g VS for food waste, agricultural residues, sugar beet bagasse, olive pulp and dairy manure respectively. The methane content of the biogas varied between 72 and 60 %. The volatile solids conversion rate for food waste was 62%.

Keywords: biogas production, organic wastes, biomethane, anaerobic digestion

Procedia PDF Downloads 253

Authors: Jacob Schwartz, Maxim Durach, Aniruddha Mitra, Abbas Rashidi, Glen Sage, Atin Adhikari

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NIOSH (National Institute for Occupational Safety and Health) approved N95 respirators are commonly used by workers in construction sites where there is a large amount of dust being produced from sawing, grinding, blasting, welding, etc., both electrostatically charged and not. A significant portion of airborne particles in construction sites could be nanoparticles created beside coarse particles. The penetration of the particles through the masks may differ depending on the size and charge of the individual particle. In field experiments relevant to this current study, we found that nanoparticles of medium size ranges are penetrating more frequently than nanoparticles of smaller and larger sizes. For example, penetration percentages of nanoparticles of 11.5 – 27.4 nm into a sealed N95 respirator on a manikin head ranged from 0.59 to 6.59%, whereas nanoparticles of 36.5 – 86.6 nm ranged from 7.34 to 16.04%. The possible causes behind this increased penetration of mid-size nanoparticles through mask filters are not yet explored. The objective of this study is to identify causes behind this unusual behavior of mid-size nanoparticles. We have considered such physical factors as Boltzmann distribution of the particles in thermal equilibrium with the air, kinetic energy of the particles at impact on the mask, Stoke’s drag force, and electrostatic forces in the mask stopping the particles. When the particles collide with the mask, only the particles that have enough kinetic energy to overcome the energy loss due to the electrostatic forces and the Stokes’ drag in the mask can pass through the mask. To understand this process, the following assumptions were made: (1) the effect of Stoke’s drag depends on the particles’ velocity at entry into the mask; (2) the electrostatic force is proportional to the charge on the particles, which in turn is proportional to the surface area of the particles; (3) the general dependence on electrostatic charge and thickness means that for stronger electrostatic resistance in the masks and thicker the masks’ fiber layers the penetration of particles is reduced, which is a sensible conclusion. In sampling situations where one mask was soaked in alcohol eliminating electrostatic interaction the penetration was much larger in the mid-range than the same mask with electrostatic interaction. The smaller nanoparticles showed almost zero penetration most likely because of the small kinetic energy, while the larger sized nanoparticles showed almost negligible penetration most likely due to the interaction of the particle with its own drag force. If there is no electrostatic force the fraction for larger particles grows. But if the electrostatic force is added the fraction for larger particles goes down, so diminished penetration for larger particles should be due to increased electrostatic repulsion, may be due to increased surface area and therefore larger charge on average. We have also explored the effect of ambient temperature on nanoparticle penetrations and determined that the dependence of the penetration of particles on the temperature is weak in the range of temperatures in the measurements 37-42°C, since the factor changes in the range from 3.17 10-3K-1 to 3.22 10-3K-1.

Keywords: respiratory protection, industrial hygiene, aerosol, electrostatic force

Procedia PDF Downloads 171

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

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

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

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Authors: Hamza Javar Magnier, Leslie V. Woodcock

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We report extensive molecular dynamics (MD) computational investigations into the thermodynamic description of supercritical properties for a model fluid that is the simplest realistic representation of atoms or molecules. The pair potential is a hard-sphere repulsion of diameter σ with a very short attraction of length λσ. When λ = 1.005 the range is so short that the model atoms are referred to as “adhesive spheres”. Molecular dimers, trimers …etc. up to large clusters, or droplets, of many adhesive-sphere atoms are unambiguously defined. This then defines percolation transitions at the molecular level that bound the existence of gas and liquid phases at supercritical temperatures, and which define the existence of a supercritical mesophase. Both liquid and gas phases are seen to terminate at the loci of percolation transitions, and below a second characteristic temperature (Tc2) are separated by the supercritical mesophase. An analysis of the distribution of clusters in gas, meso- and liquid phases confirms the colloidal nature of this mesophase. The general phase behaviour is compared with both experimental properties of the water-steam supercritical region and also with formally exact cluster theory of Mayer and Mayer. Both are found to be consistent with the present findings that in this system the supercritical mesophase narrows in density with increasing T > Tc and terminates at a higher Tc2 at a confluence of the primary percolation loci. The expended plot of the MD data points in the mesophase of 7 critical and supercritical isotherms in highlight this narrowing in density of the linear-slope region of the mesophase as temperature is increased above the critical. This linearity in the mesophase implies the existence of a linear combination rule between gas and liquid which is an extension of the Lever rule in the subcritical region, and can be used to obtain critical parameters without resorting to experimental data in the two-phase region. Using this combination rule, the calculated critical parameters Tc = 0.2007 and Pc = 0.0278 are found be agree with the values found by of Largo and coworkers. The properties of this supercritical mesophase are shown to be consistent with an alternative description of the phenomenon of critical opalescence seen in the supercritical region of both molecular and colloidal-protein supercritical fluids.

Keywords: critical opalescence, supercritical, square-well, percolation transition, critical parameters.

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Authors: Rossy G. Olan Villegas, Gerardo Acosta Garcia, Aurea Bernardino Nicanor, Leopoldo Gonzalez Cruz, Humberto Ramirez Medina

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Capsicum annuum is a crop of agricultural and economic importance in Mexico and other countries. The fruit (pepper) contains bioactive components such as carotenoids, phenolic compounds and capsaicinoids that improve health. However, pepper cultivation is affected by biotic and abiotic factors that decrease yield. Some phytohormones like gibberellins and auxins induce the formation and development of fruit in several plants. In this study, we evaluated the effect of the exogenous application of phytohormones like gibberellic acid and indolbutyric acid on fruit development of jalapeno pepper plants, the protein profile of plant tissues, the accumulation of bioactive compounds and antioxidant activity in the pericarp and seeds. For that, plants were sprinkled with these phytohormones. The fruit collection for the control, indolbutyric acid and gibberellic acid treatments was 7 peppers per plant; however, for the treatment that combines indolbutyric acid and gibberellic acid, a fruit with the shortest length (1.52 ± 1.00 cm) and weight (0.41 ± 1.0 g) was collected compared to fruits of plants grown under other treatments. The length (4,179 ± 0,130 cm) and weight of the fruit (8,949 ± 0.583 g) increased in plants treated with indolbutyric acid, but these characteristics decreased with the application of GA3 (length of 3,349 ± 0.127 cm and a weight 4,429 ± 0.144 g). The content of carotenes and phenolic compounds increased in plants treated with GA3 (1,733 ± 0.092 and 1,449 ± 0.009 mg / g, respectively) or indolbutyric acid (1,164 ± 0.042 and 0.970 ± 0.003 mg / g). However, this effect was not observed in plants treated with both phytohormones (0.238 ± 0.021 and 0.218 ± 0.004 mg / g). Capsaicin content was higher in all treatments; but it was more noticeable in plants treated with both phytohormones, the value being 0.913 ± 0.001 mg / g (three times greater in amount). The antioxidant activity was measured by 3 different assays, 2,2-diphenyl-1-picrylhydrazyl (DPPH), antioxidant power of ferric reduction (FRAP) and 2,2'-Azinobis-3-ethyl-benzothiazoline-6-sulfonic acid ( ABTS) to find the minimum inhibitory concentration of the reducing radical (IC50 and EC50). Significant differences were observed from the application of the phytohormone, being the fruits treated with gibberellins, which had a greater accumulation of bioactive compounds. Our results suggest that the application of phytohormones modifies the development of fruit and its content of bioactive compounds.

Keywords: auxins, capsaicinoids, carotenoids, gibberellins

Procedia PDF Downloads 91

Authors: Menna Ewida, Dalal Abou El-Ella, Dina Lasheen, Huessin El-Subbagh

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Introduction: Vascular Endothelial Growth Factor receptor (VEGF) is a signal protein produced by cells that stimulates vasculogenesis to create new blood vessels. VEGF family binds to three trans-membrane tyrosine kinase receptors,Dihydrofolate reductase (DHFR) is an enzyme of crucial importance in medicinal chemistry. DHFR catalyzes the reduction 7,8 dihydro-folate to tetrahydrofolate and intimately couples with thymidylate synthase which is a pivotal enzyme that catalysis the reductive methylation of deoxyuridine monophosphate (dUMP) to deoxythymidine monophosphate (dTMP) utilizing N5,N10-methylene tetrahydrofolate as a cofactor which functions as the source of the methyl group. Purpose: Novel substituted Thiazole agents were designed as DHFR and VEGF-TK inhibitors with increased synergistic activity and decreased side effects. Methods: Five series of compounds were designed with a rational that mimic the pharmacophoric features present in the reported active compounds that target DHFR & VEGFR. These molecules were docked against Methotrexate & Sorafenib as controls. An in silico ADMET study was also performed to validate the bioavailability of the newly designed compounds. The in silico molecular docking & ADMET study were also applied to the non-classical antifolates for comparison. The interaction energy comparable to that of MTX for DHFRI and Sorafenib for VEGF-TKI activity were recorded. Results: Compound 5 exhibited the highest interaction energy when docked against Sorafenib, While Compound 9 showed the highest interaction energy when docked against MTX with the perfect binding mode. Comparable results were also obtained for the ADMET study. Most of the compounds showed absorption within (95-99) zone which varies according to the type of substituents. Conclusions: The Substituted Thiazole Analogues could be a suitable template for antitumor drugs that possess enhanced bioavailability and act as DHFR and VEGF-TK inhibitors.

Keywords: anti-tumor agents, DHFR, drug design, molecular modeling, VEGFR-TKIs

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Authors: Min Liu, Jirui Gong, Qinpu Luo, Lili Yang, Bo Yang, Zihe Zhang, Yan Pan, Zhanwei Zhai

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Grazing disturbance is one of the important land-use types that affect plant growth and ecosystem processes. In order to study the responses of dominant species to grazing in the semiarid temperate grassland of Inner Mongolia, we set five grazing intensity plots: a control and four levels of grazing (light (LG), moderate (MG), heavy (HG) and extreme heavy grazing (EHG)) to test the morphological and physiological responses of Stipa grandis, Leymus chinensis at the individual levels. With the increase of grazing intensity, Stipa grandis and Leymus chinensis both exhibited reduced plant height, leaf area, stem length and aboveground biomass, showing a significant dwarf phenomenon especially in HG and EHG plots. The photosynthetic capacity decreased along the grazing gradient. Especially in the MG plot, the two dominant species have lowest net photosynthetic rate (Pn) and water use efficiency (WUE). However, in the HG and EHG plots, the two species had high light saturation point (LSP) and low light compensation point (LCP), indicating they have high light-use efficiency. They showed a stimulation of compensatory photosynthesis to the remnant leaves as compared with grasses in MG plot. For Leymus chinensis, the lipid peroxidation level did not increase with the low malondialdehyde (MDA) content even in the EHG plot. It may be due to the high enzymes activity of superoxide dismutase (SOD) and peroxidase (POD) to reduce the damage of reactive oxygen species. Meanwhile, more carbohydrate was stored in the leaf of Leymus chinensis to provide energy to the plant regrowth. On the contrary, Stipa grandis showed the high level of lipid peroxidation especially in the HG and EHG plots with decreased antioxidant enzymes activity. The soluble protein content did not change significantly in the different plots. Therefore, with the increase of grazing intensity, plants changed morphological and physiological traits to defend themselves effectively to herbivores. Leymus chinensis is more resistant to grazing than Stipa grandis in terms of tolerance traits, particularly under heavy grazing pressure.

Keywords: antioxidant enzymes activity, grazing density, morphological responses, photosynthesis

Procedia PDF Downloads 337

Authors: O. A. Adebiyi, A. O. Oni, A. O. K. Adeshehinwa, I. O. Adejumo

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In forty nine (49) days, twenty four (24) growing pigs (Landrace x Large white) with an average weight of 17 ±2.1kg were allocated to four experimental treatments T1 (dry mash without probiotics), T2 (wet feed without probiotics), T3 (dry mash + Saccaromyces cereviseae probiotics) and T4 (wet feed + Saccaromyces cereviseae probiotics) which were replicated three times with two pigs per replicate in a completely randomised design. The basal feed (dry feed) was formulated to meet the nutritional requirement of the animal with crude protein of 18.00% and metabolisable energy of 2784.00kcal/kgME. Growth pattern, faecal microbial load and behavioural activities (eating, drinking, physical pen interaction and frequency of visiting the drinking troughs) were accessed. Pigs fed dry mash without probiotics (T1) had the highest daily feed intake among the experimental animals (1.10kg) while pigs on supplemented diets (T3 and T4) had an average daily feed intake of 0.95kg. However, the feed conversion ratio was significantly (p < 0.05) affected with pigs on T3 having least value of 6.26 compared those on T4 (wet feed + Saccaromyces cereviseae) with means of 7.41. Total organism counts varied significantly (p < 0.05) with pigs on T1, T2, T3 and T4 with mean values of 179.50 x106cfu; 132.00 x 106cfu; 32.00 x 106cfu and 64.50 x 106cfu respectively. Coliform count was also significantly (p < 0.05) different among the treatments with corresponding values of 117.50 x 106cfu; 49.00 x 106cfu, 8.00 x 106cfu for pigs in T1, T2 and T4 respectively. The faecal Saccaromyces cereviseae was significantly lower in pigs fed supplemented diets compared to their counterparts on unsupplemented diets. This could be due to the inability of yeast organisms to be voided easily through feaces. The pigs in T1 spent the most time eating (7.88%) while their counterparts on T3 spent the least time eating. The corresponding physical pen interaction times expressed in percentage of a day for pigs in T1, T2, T3 and T4 are 6.22%, 5.92%, 4.04% and 4.80% respectively. These behavioural responses exhibited by these pigs (T3) showed that little amount of dry feed supplemented with probiotics is needed for better performance. The water intake increases as a result of the dryness of the feed with consequent decrease in pen interaction and more time was spent resting than engaging in other possible vice-habit like fighting or tail biting. Pigs fed dry feed (T3) which was supplemented with Saccaromyces cereviseae probiotics had a better overall performance, least faecal microbial load than wet fed pigs either supplemented with Saccaromyces cereviseae or non-supplemented.

Keywords: behaviour, feed forms, feed utilization, growth, microbial

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Authors: Manal Elsayed Elkheshin, Rasha Saleh Mansour, Mohamed Fawzy Mohamed Eldnasury, Mamdouh Elbadry Mohamed

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This research Aims to identify the main factors affecting the production and the fish consumption in Egypt, through the econometric estimation for various forms functions of fish production and fish consumption during the period (1991-2014), as the aim of this research to forecast the production and the fish consumption in Egypt until 2020, through determine the best standard methods using (ARIMA).This research also aims to the economic feasibility of the production of fish in aquaculture farms study; investment cost and represents the value of land, buildings, equipment and irrigation. Aquaculture requires three types of fish (Tilapia, carp fish, and mullet fish), and the total area of the farm, about an acre. The annual Fish production from this project about 3.5 tons. The annual investment costs of about 50500 pounds, Find conclude that the project can repay the cost of their investments after about 4 years and 5 months, and therefore recommend the implementation of the project, and internal rate of return reached (IRR) of about 22.1%, where it is clear that the rate of large internal rate of return, and achieves pound invested in this project annual return is estimated at 22.1 pounds, more than the opportunity cost, so we recommend the need to implement the project.Recommendations:1. Increasing the fish agriculture to decrease the gap of animal protein. 2.Increasing the number of mechanism fishing boats, and the provision of transport equipped to maintain the quality of fish production. 3.Encourage and attract the local and foreign investments, providing advice to the investor on the aquaculture field. 4. Action newsletters awareness of the importance of these projects where these projects resulted in a net profit after recovery in less than five years, IRR amounted to about 23%, which is much more than the opportunity cost of a bank interest rate is about 7%, helping to create work and graduates opportunities, and contribute to the reduction of imports of the fish, and improve the performance of the food trade balance.

Keywords: equation model, individual share, red meat, consumption, production, endogenous variable, exogenous variable, financial performance evaluates fish culture, feasibility study, fish production, aquaculture

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Authors: Francesca Lombardi, Francesca Rosaria Augello, Benedetta Cinque, Maria Grazia Cifone, Paola Palumbo

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Glioblastoma multiforme (GBM) is a high-grade primary brain tumor refractory to current forms of treatment. The survival benefits of patients with GBM remain unsatisfactory due to the intrinsic or acquired resistance to temozolomide (TMZ), an alkylating agent, used as the first-line chemotherapeutic drug to treat GBM patients. Its cytotoxic effect is visualized by the induction of O6-methylguanine (O6MeG) within DNA. Cyclooxygenase-2 (COX-2), an inflammation-associated enzyme, has been implicated in tumorigenesis and progression of GBM, its inhibition shows anticancer activities. In the present study, it was verified if the combination of a COX-2 selective inhibitor, NS398, with TMZ could counteract the TMZ resistance. In particular, the effect of NS398 mixed with TMZ was investigated in the GBM TMZ-resistant cell line, T98G. Cells were pretreated with NS398 (100µM, 24 hours) and then exposed to TMZ alone (200µM), NS398 alone, or both for 72 hours, after which cell growth rate and cycle phases, as well as apoptosis level, were evaluated. Coadministration of NS398 and TMZ caused a significant decrease in cell growth and a progressive increase of dead cells detected by trypan blue staining. Moreover, a significant level of apoptotic cell percentage and alteration of cell cycle phases were observed in T98G treated with TMZ-NS398 combination when compared to untreated cells or TMZ-treated cells. TMZ-resistant tumors, as GBM, express elevated levels of DNA repair enzyme O6-methylguanine-DNA methyltransferase (MGMT). The mixture drastically reduced MGMT expression in the TMZ-resistant cell line T98G, known to express high levels of MGMT basically. Moreover, while TMZ alone did not influence the COX-2 protein expression, the combination successfully reduced it. In conclusion, these results demonstrated that NS398 enhanced the efficacy of TMZ through cell number reduction, apoptosis induction, and decreased MGMT levels, suggesting the ability of drug combination to reduce the chemoresistance. This drug combination deserves attention and could be considered as a promising therapeutic strategy for GBM patients.

Keywords: COX-2, COX-2 inhibitor, glioblastoma, NS398, T98G, temozolomide

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Authors: Delgado-Meza M., Minor-Pérez H.

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Protease is the generic name of enzymes that hydrolyze proteins. These are classified in the subgroup EC3.4.11-99X of the classification enzymes. In food technology the proteolysis is used to modify functional and nutritional properties of food, and in some cases this proteolysis may cause food spoilage. In general, seafood and rainbow trout have accelerated decomposition process once it has done its capture, due to various factors such as the endogenous enzymatic activity that can result in loss of structure, shape and firmness, besides the release of amino acid precursors of biogenic amines. Some studies suggest the use of protease inhibitors from legume as biological regulators of proteolytic activity. The enzyme inhibitors are any substance that reduces the rate of a reaction catalyzed by an enzyme. The objective of this study was to evaluate the reduction of the proteolytic activity of enzymes in extracts of rainbow trout with protease inhibitors obtained from chickpea flour. Different proportions of rainbow trout enzyme extract (75%, 50% and 25%) and extract chickpea enzyme inhibitors were evaluated. Chickpea inhibitors were obtained by mixing 5 g of flour in 30 mL of pH 7.0 phosphate buffer. The sample was centrifuged at 8000 rpm for 10 min. The supernatant was stored at -15°C. Likewise, 20 g of rainbow trout were ground in 20 mL of phosphate buffer solution at pH 7.0 and the mixture was centrifuged at 5000 rpm for 20 min. The supernatant was used for the study. In each treatment was determined the specific enzymatic activity with the technique of Kunitz, using hemoglobin as substrate for the enzymes acid fraction and casein for basic enzymes. Also biuret protein was quantified for each treatment. The results showed for fraction of basic enzymes in the treatments evaluated, that were inhibition of endogenous enzymatic activity. Inhibition values compared to control were 51.05%, 56.59% and 59.29% when the proportions of endogenous enzymes extract rainbow trout were 75%, 50% and 25% and the remaining volume used was extract with inhibitors. Treatments with acid enzymes showed no reduction in enzyme activity. In conclusion chickpea flour reduced the endogenous enzymatic activity of rainbow trout, which may favor its application to increase the half-life of this food. The authors acknowledge the funding provided by the CONACYT for the project 131998.

Keywords: rainbouw trout, enzyme inhibitors, proteolysis, enzyme activity

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Authors: Rosa Figueroa, Christopher Flores

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Text categorization is the problem of assigning a new document to a set of predetermined categories, on the basis of a training set of free-text data that contains documents whose category membership is known. To train a classification model, it is necessary to extract characteristics in the form of tokens that facilitate the learning and classification process. In text categorization, the feature extraction process involves the use of word sequences also known as N-grams. In general, it is expected that documents belonging to the same category share similar features. The Smith-Waterman (SW) algorithm is a dynamic programming algorithm that performs a local sequence alignment in order to determine similar regions between two strings or protein sequences. This work explores the use of SW algorithm as an alternative to feature extraction in text categorization. The dataset used for this purpose, contains 2,610 annotated documents with the classes Obese/Non-Obese. This dataset was represented in a matrix form using the Bag of Word approach. The score selected to represent the occurrence of the tokens in each document was the term frequency-inverse document frequency (TF-IDF). In order to extract features for classification, four experiments were conducted: the first experiment used SW to extract features, the second one used unigrams (single word), the third one used bigrams (two word sequence) and the last experiment used a combination of unigrams and bigrams to extract features for classification. To test the effectiveness of the extracted feature set for the four experiments, a Support Vector Machine (SVM) classifier was tuned using 20% of the dataset. The remaining 80% of the dataset together with 5-Fold Cross Validation were used to evaluate and compare the performance of the four experiments of feature extraction. Results from the tuning process suggest that SW performs better than the N-gram based feature extraction. These results were confirmed by using the remaining 80% of the dataset, where SW performed the best (accuracy = 97.10%, weighted average F-measure = 97.07%). The second best was obtained by the combination of unigrams-bigrams (accuracy = 96.04, weighted average F-measure = 95.97) closely followed by the bigrams (accuracy = 94.56%, weighted average F-measure = 94.46%) and finally unigrams (accuracy = 92.96%, weighted average F-measure = 92.90%).

Keywords: comorbidities, machine learning, obesity, Smith-Waterman algorithm

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Authors: Yee-Seul So, Guiseppe Ianiri, Alex Idnurm, Yong-Sun Bahn

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Tor1 is a serine/threonine protein kinase that is widely conserved across eukaryotic species. Tor1 was first identified in Saccharomyces cerevisiae as a target of rapamycin (TOR). The TOR pathway has been implicated in regulating cellular responses to nutrients, proliferation, translation, transcription, autophagy, and ribosome biogenesis. Here we identified two homologues of S. cerevisiae Tor proteins, CNAG_06642 (Tor1) and CNAG_05220 (Tlk1, TOR-like kinase 1), in Cryptococcus neoformans causing a life-threatening fungal meningoencephalitis. Both Tor1 and Tlk1 have rapamycin-binding (RB) domains but Tlk1 has truncated RB form. To study the TOR-signaling pathway in the fungal pathogen, we attempt to construct the tor1Δ and tlk1Δ mutants and phenotypically analyze them. Although we failed to construct the tor1Δ mutant, we successfully construct the tlk1Δ mutant. The tlk1Δ mutant does not exhibit any discernable phenotypes, suggesting that Tlk1 is dispensable in C. neoformans. The essentiality of TOR1 is independently confirmed by constructing the TOR1 promoter replacement strain by using a copper transporter 4 (CTR4) promoter and the TOR1/tor1 heterozygous mutant in diploid C. neoformans strain background followed by sporulation analysis. To further analyze the function of Tor1, we construct TOR1 overexpression mutant using a constitutively active histone H3 in C. neoformans. We find that the Tor1 overexpression mutant is resistant to rapamycin but the tlk1Δ mutant does not exhibit any altered resistance to rapamycin, further confirming that Tor1, but not Tlk1, is critical for TOR signaling. Furthermore, we found that Tor1 is involved in response to diverse stresses, including genotoxic stress, oxidative stress, thermo-stress, antifungal drug treatment, and production of melanin. To identify any TOR-related transcription factors, we screened C. neoformans transcription factor library that we constructed in our previous study and identified several potential downstream factors of Tor1, including Atf1, Crg1 and Bzp3. In conclusion, the current study provides insight into the role of the TOR signaling pathway in human fungal pathogens as well as C. neoformans.

Keywords: fungal pathogen, serine/threonine kinase, target of rapamycin, transcription factor

Procedia PDF Downloads 200

Authors: Hamdy Khamis Korayem, Manal Yehia Tayel, Abeer Shawky El Hadedy, Emmanuel Kamal Aziz Saba, Shimaa Badr Abdelnaby Badr

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The aim of the current study was to demonstrate whether serum Interleukin-18 (IL-18) is increased in rheumatoid arthritis (RA) and its correlation with disease activity, functional disability and quality of life in RA patients. The study included 30 RA patients and 20 healthy normal control subjects. The RA patients were diagnosed according to the 2010 ACR/EULAR classification criteria for RA with the exclusion of those who had diabetes mellitus, endocrine disorders, associated rheumatologic diseases, viral hepatitis B or C and other diseases with increased serum IL-18 level. All patients were subjected to clinical evaluation of the musculoskeletal system. Disease activity was assessed by disease activity score 28 with 4 variables (DAS 28). Functional disability was assessed by health assessment questionnaire disability index (HAQ-DI). The quality of life was assessed by Short form-36 (SF-36) questionnaire. Radiological assessment of both hands and feet by Sharp/van der Heijde (SvH) scoring method. Laboratory parameters including erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibody (ACPA) were assessed in patients and serum level of IL-18 in both patients and control subjects. There was no statistically significant difference between patient and control group as regards age and sex. Among patients, 29 % were females and the age range was between 25 to 55 years. Extra-articular manifestations were presented in 56.7% of the patients. The mean of DAS 28 score was 5.73±1.46 and that of HAQ-DI was 1.22±0.72 while that of SF-36 was 40.03±13.96. The level of serum IL-18 was significantly higher in patients than in the control subjects (P= 0.030). Serum IL-18 was correlated with ACPA among the patient group. There were no statistically significant correlations between serum IL-18 and DAS28, HAQ-DI, SF-36, total SvH score and the other laboratory results. In conclusion, IL-18 is significantly higher in RA patient than in healthy control subjects and positively correlated with ACPA level. IL-18 is associated with extra-articular manifestations. However, it is not correlated with other laboratory parameters, disease activity, functional disability, quality of life nor radiological severity.

Keywords: disease activity score, Interleukin-18, quality of life assessment, rheumatoid arthritis

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Authors: Iwona Cieślik, Władysław Migdał, Kinga Topolska, Ewa Cieślik

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The consumption of fish is recommended as a means of preventing serious diseases, especially cardiovascular problems. Fish is known to be a valuable source of protein (rich in essential amino acids), unsaturated fatty acids, fat-soluble vitamins, macro- and microelements. However, it can also contain several contaminants (e.g. pesticides, heavy metals) that may pose considerable risks for humans. Among others, pesticide are of special concern. Their widespread use has resulted in the contamination of environmental compartments, including water. The occurrence of pesticides in the environment is a serious problem, due to their potential toxicity. Therefore, a systematic monitoring is needed. The aim of the study was to determine the organochlorine and organophosphate pesticide residues in fish muscle tissues of the pike (Esox lucius, L.) and the rainbow trout (Oncorhynchus mykkis, Walbaum) by a modified QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) method, using Gas Chromatography Quadrupole Mass Spectrometry (GC/Q-MS), working in selected-ion monitoring (SIM) mode. The analysis of α-HCH, β-HCH, lindane, diazinon, disulfoton, δ-HCH, methyl parathion, heptachlor, malathion, aldrin, parathion, heptachlor epoxide, γ-chlordane, endosulfan, α-chlordane, o,p'-DDE, dieldrin, endrin, 4,4'-DDD, ethion, endrin aldehyde, endosulfan sulfate, 4,4'-DDT, and metoxychlor was performed in the samples collected in the Carp Valley (Malopolska region, Poland). The age of the pike (n=6) was 3 years and its weight was 2-3 kg, while the age of the rainbow trout (n=6) was 0.5 year and its weight was 0.5-1.0 kg. Detectable pesticide (HCH isomers, endosulfan isomers, DDT and its metabolites as well as metoxychlor) residues were present in fish samples. However, all these compounds were below the limit of quantification (LOQ). The other examined pesticide residues were below the limit of detection (LOD). Therefore, the levels of contamination were - in all cases - below the default Maximum Residue Levels (MRLs), established by Regulation (EC) No 396/2005 of the European Parliament and of the Council. The monitoring of pesticide residues content in fish is required to minimize potential adverse effects on the environment and human exposure to these contaminants.

Keywords: contaminants, fish, pesticides residues, QuEChERS method

Procedia PDF Downloads 192

Authors: Amna Shoaib, Sidrah Hanif, Rashid Mehmood

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Current research work was carried out to check influence of farmyard manure (FYM) in Lycopersicon esculentum L. against Fusarium oxysporum f. sp. lycopersici (FO) in copper polluted soil. Silt-loam soil naturally enriched with 70 ppm of Cu was inoculated with 1 x 106 spore suspensions of FO and incorporated with 0%, 1%, 1.5% or 2% FYM. The multilateral interaction of host-pathogen-metal-organic amendment was assessed in terms of morphology, growth, yield, physiology, biochemistry and metal uptake in tomato plant after 30 and 60 days of sowing. When soil was inoculated with FO, plant growth and biomass were significantly increased during vegetative stage, while declining during flowering stage with substantial increase in productivity over control. Infected plants exhibited late wilting and disease severity was found on 26-50% of plant during reproductive stage. Incorporation of up to 1% FYM suppressed disease severity, improved plant growth and biomass, while it decreased yield. Rest of manure doses was found ineffective in suppressing disease. Content of total chlorophyll, sugar and protein were significantly declined in FO inoculated plants and incorporation of FYM caused significant reduction or no influence on sugar and chlorophyll content, and no pronounced difference among different FYM doses were observed. On the other hand, proline, peroxidase, catalase and nitrate reductase activity were found to be increased in infected plants and incorporation of 1-2% FYM further enhanced the activity of these enzymes. Tomato plant uptake of 30-40% of copper naturally present in the soil and incorporation of 1-2% FYM markedly decreased plant uptake of metal by 15-30%, while increased Cu retention in soil. Present study concludes that lower dose (1%) of FYM could be used to manage disease, increase growth and biomass, while being ineffective for yield and productivity in Cu-polluted soil. Altered physiology/biochemistry of plant in response to any treatment could be served as basis for resistant against pathogen and metal homeostasis in plants.

Keywords: Lycopersicon esculentum, copper, Fusarium wilt, farm yard manure

Procedia PDF Downloads 388

Authors: Maha Z. Rizk, Sami A. Fattah, Heba M. Darwish, Sanaa A. Ali, Mai O. Kadry

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Although it is known that nano-TiO2 and other nanoparticles can induce liver toxicity, the mechanisms and the molecular pathogenesis are still unclear. The present study investigated some biochemical indices of nano-sized Titanium dioxide (TiO2 NPS) toxicity in mice liver and the ameliorative efficacy of individual and combined doses of idebenone, carnosine and vitamin E. Nano-anatase TiO2 (21 nm) was administered as a total oral dose of 2.2 gm/Kg daily for 2 weeks followed by the afore-mentioned antioxidants daily either individually or in combination for 1month. TiO2-NPS induced a significant elevation in serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and hepatic oxidative stress biomarkers [lipid peroxides (LP), and nitric oxide levels (NOX), while it significantly reduced glutathione reductase (GR), reduced glutathione (GSH) and glutathione peroxidase(GPX) levels. Moreover the quantitative RT-PCR analysis showed that nano-anatase TiO2 can significantly alter the mRNA and protein expressions of the fibrotic factors TGF-B1, VEGFand Smad-2. Histopathological examination of hepatic tissue reinforced the previous biochemical results. Our results also implied that inflammatory responses and liver injury may be involved in nano-anatase TiO2-induced liver toxicity Tumor necrosis factor-α (TNF-α) and Interleukin -6 (IL-6) and increased the percent of DNA damage which was assessed by COMET assay in addition to the apoptotic marker Caspase-3. Moreover mRNA gene expression observed by RT-PCR showed a significant overexpression in nuclear factor relation -2 (Nrf2), nuclear factor kappa beta (NF-Kβ) and the apoptotic factor (bax), and a significant down regulation in the antiapoptotic factor (bcl2) level. In conclusion idebenone, carnosine and vitamin E ameliorated the deviated previously mentioned parameters with variable degrees with the most pronounced role in alleviating the hazardous effect of TiO2 NPS toxicity following the combination regimen.

Keywords: Nano-anatase TiO2, TGF-B1, SMAD-2

Procedia PDF Downloads 405

Authors: Jienny Lee, In-Soo Cho, Sang-Ho Cha

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Adult mesenchymal stem cells (MSCs) have been investigated using preclinical approaches for tissue regeneration. Porcine MSCs (pMSCs) are capable of growing and attaching to plastic with a fibroblast-like morphology and then differentiating into bone, adipose, and cartilage tissues in vitro. This study was conducted to investigate the proliferating abilities, differentiation potentials, and multipotency of miniature pig adipose tissue-derived MSCs (mpAD-MSCs) with or without long-term cryopreservation, considering that cryostorage has the potential for use in clinical applications. After confirming the characteristics of the mpAD-MSCs, we examined the effect of long-term cryopreservation (> 2 years) on expression of cell surface markers (CD34, CD90 and CD105), proliferating abilities (cumulative population doubling level, doubling time, colony-forming unit, and MTT assay) and differentiation potentials into mesodermal cell lineages. As a result, the expression of cell surface markers is similar between thawed and fresh mpAD-MSCs. However, long-term cryopreservation significantly lowered the differentiation potentials (adipogenic, chondrogenic, and osteogenic) of mpAD-MSCs. When compared with fresh mpAD-MSCs, thawed mpAD-MSCs exhibited lower expression of mesodermal cell lineage-related genes such as peroxisome proliferator-activated receptor-g2, lipoprotein lipase, collagen Type II alpha 1, osteonectin, and osteocalcin. Interestingly, long-term cryostoraged mpAD-MSCs exhibited significantly higher cell viability than the fresh mpAD-MSCs. Long-term cryopreservation induced a 30% increase in the cell viability of mpAD-MSCs when compared with the fresh mpAD-MSCs at 5 days after thawing. However, long-term cryopreservation significantly lowered expression of stemness markers such as Oct3/4, Sox2, and Nanog. Furthermore, long-term cryopreservation negatively affected expression of senescence-associated genes such as telomerase reverse transcriptase and heat shock protein 90 of mpAD-MSCs when compared with the fresh mpAD-MSCs. The results from this study might be important for the successful application of MSCs in clinical trials after long-term cryopreservation.

Keywords: mesenchymal stem cells, cryopreservation, stemness, senescence

Procedia PDF Downloads 210

Authors: Pantea Salehizadeh, Martin P. Bucknall, Robert Driscoll, Jayashree Arcot, George Srzednicki

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Banana is one of the most important crops produced in large quantities in tropical and sub-tropical countries. Of the total plant material grown, approximately 40% is considered waste and left in the field to decay. This practice allows fungal diseases such as Sigatoka Leaf Spot to develop, limiting plant growth and spreading spores in the air that can cause respiratory problems in the surrounding population. The pseudostem is considered a waste residue of production (60 to 80 tonnes/ha/year), although it is a good source of dietary fiber and volatile organic compounds (VOC’s). Strategies to process banana pseudostem into palatable, nutritious and marketable food materials could provide significant social and economic benefits. Extraction of VOC’s with desirable odor from dried and fresh pseudostem could improve the smell of products from the confectionary and bakery industries. Incorporation of banana pseudostem flour into bakery products could provide cost savings and improve nutritional value. The aim of this study was to determine the effects of drying methods and different banana species on the profile of volatile aroma compounds in dried banana pseudostem. The banana species analyzed were Musa acuminata and Musa balbisiana. Fresh banana pseudostem samples were processed by either freeze-drying (FD) or heat pump drying (HPD). The extraction of VOC’s was performed at ambient temperature using vacuum distillation and the resulting, mostly aqueous, distillates were analyzed using headspace solid phase microextraction (SPME) gas chromatography – mass spectrometry (GC-MS). Optimal SPME adsorption conditions were 50 °C for 60 min using a Supelco 65 μm PDMS/DVB Stableflex fiber1. Compounds were identified by comparison of their electron impact mass spectra with those from the Wiley 9 / NIST 2011 combined mass spectral library. The results showed that the two species have notably different VOC profiles. Both species contained VOC’s that have been established in literature to have pleasant appetizing aromas. These included l-Menthone, D-Limonene, trans-linlool oxide, 1-Nonanol, CIS 6 Nonen-1ol, 2,6 Nonadien-1-ol, Benzenemethanol, 4-methyl, 1-Butanol, 3-methyl, hexanal, 1-Propanol, 2-methyl- acid، 2-Methyl-2-butanol. Results show banana pseudostem VOC’s are better preserved by FD than by HPD. This study is still in progress and should lead to the optimization of processing techniques that would promote the utilization of banana pseudostem in the food industry.

Keywords: heat pump drying, freeze drying, SPME, vacuum distillation, VOC analysis

Procedia PDF Downloads 298

Authors: Djeddi Khaled, Houssou Hind, Miloudi Abdellatif, Rabah Siham

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In the field of veterinary medicine, there is a growing application of artificial intelligence (AI) for diagnosing bovine mastitis, a prevalent inflammatory disease in dairy cattle. AI technologies, such as automated milking systems, have streamlined the assessment of key metrics crucial for managing cow health during milking and identifying prevalent diseases, including mastitis. These automated milking systems empower farmers to implement automatic mastitis detection by analyzing indicators like milk yield, electrical conductivity, fat, protein, lactose, blood content in the milk, and milk flow rate. Furthermore, reports highlight the integration of somatic cell count (SCC), thermal infrared thermography, and diverse systems utilizing statistical models and machine learning techniques, including artificial neural networks, to enhance the overall efficiency and accuracy of mastitis detection. According to a review of 15 publications, machine learning technology can predict the risk and detect mastitis in cattle with an accuracy ranging from 87.62% to 98.10% and sensitivity and specificity ranging from 84.62% to 99.4% and 81.25% to 98.8%, respectively. Additionally, machine learning algorithms and microarray meta-analysis are utilized to identify mastitis genes in dairy cattle, providing insights into the underlying functional modules of mastitis disease. Moreover, AI applications can assist in developing predictive models that anticipate the likelihood of mastitis outbreaks based on factors such as environmental conditions, herd management practices, and animal health history. This proactive approach supports farmers in implementing preventive measures and optimizing herd health. By harnessing the power of artificial intelligence, the diagnosis of bovine mastitis can be significantly improved, enabling more effective management strategies and ultimately enhancing the health and productivity of dairy cattle. The integration of artificial intelligence presents valuable opportunities for the precise and early detection of mastitis, providing substantial benefits to the dairy industry.

Keywords: artificial insemination, automatic milking system, cattle, machine learning, mastitis

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Authors: Mohammad Nasb, Muhammad Rehan, Chen Hong

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Background Meniscus defects critically alter knee function and lead to degenerative changes. Regenerative medicine applications including stem cell transplantation have showed a promising efficacy in finding alternatives to overcome traditional treatment limitations. However, stem cell therapy remains limited due to the substantially reduced viability and inhibitory microenvironment. Since tissue growth and repair are under the control of biochemical and mechanical signals, several approaches have recently been investigated (e.g., low intensity pulsed ultrasound [LIPUS]) to promote the regeneration process. This study employed LIPUS to improve growth and osteogenic differentiation of mesenchymal stem cells derived from human embryonic stem cells to improve the regeneration of meniscus tissue. Methodology: The Mesenchymal stromal cells (MSCs) were transplanted into the epicenter of the injured meniscus in rabbits, which were randomized into two main groups: a treatment group (n=32 New Zealand rabbits) including 4 subgroups of 8 rabbits in each subgroup (LIPUS treatment, MSC treatment, LIPUS with MSC and control), and a second group (n=9) to track implanted cells and their progeny using green fluorescence protein (GFP). GFP consists of the MSC and LIPUS-MSC combination subgroups. Rabbits were then subjected to histological, immunohistochemistry, and MRI assessment. Results: The quantity of the newly regenerated tissue in the combination treatment group that had Ultrasound irradiation after mesenchymal stem cells were better at all end points. Likewise, Tissue quality scores were also greater in knees treated with both approaches compared with controls and single treatment at all end points, achieving significance at twelve and twenty-four weeks [p < 0.05], and [p = 0.008] at twelve weeks. Differentiation into type-I and II collagen-expressing cells were higher in the combination group at up to twenty-four weeks. Conclusions: the combination of mesenchymal stem cells and LIPUS showed greater adhering to the sites of meniscus injury, differentiate into cells resembling meniscal fibrochondrocytes, and improve both quality and quantity of meniscal regeneration.

Keywords: stem cells, regenerative medicine, osteoarthritis, knee

Procedia PDF Downloads 93

Authors: Yu-Ju Hsieh, Lin-Hung Lin, Wen-Hui Chang

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This study was designed as an interventional research and intended to validate whether the introduction of drinking vinegar every day can reduce and even prevent urinary tract infection in Taiwan home stayed disabilities who using indwelling catheter. The data was collected from the subjects who have received home care case at northern Taiwan, according to the questionnaire and a medical records retroactive methodology, the subjects were informed and consent to drink 15ml of table vinegar in a daily diet, and through routine urine testing and culture study. Home care nurses would assist collecting urine at the point of before and after a meal from total 35 studied subjects per month, and total collected 4 times for testing. The results showed that when the average age of study subjects was 65.46 years and catheter indwelling time was 15 years, drinking table vinegar could inhibit the activity of E. coli O157: H7 and reduce its breeding. Before drinking table vinegar daily, the subjects’ urine pH value was 7.0-8.0, and the average was 7.5, and the urine PH value dropped to 6.5 after drinking table vinegar for a month. There were two purple urine cases whose urine were changed from purple to normal color after two weeks of drinking, and the protein and bacteria values of urine gradually improved. Urine smell unpleasant before attending to this study, and the symptom improved significantly only after 1 week, and the urine smell returned to normal ammonia and became clean after 1 month later. None of these subjects received treatment in a hospital due to urinary tract infection, and there were no signs of bleeding in all cases during this study. The subjects of this study are chronic patients with a long-term bedridden catheterization; drinking cranberry juice is an economic burden for them, and also highly prohibited for diabetes patients. By adapting to use cheaper table vinegar to acidified urine and improve its smell and ease Purple Urine Syndrome, to furthermore, proven urinary tract infection, it can also to reduce the financial burden on families, the cost of social resources and the rate of re-admission.

Keywords: table vinegar, urinary tract infection, disability patients, long-term indwelling catheter

Procedia PDF Downloads 236

Authors: Rio Hirakawa, Christian Gundlach, Sven Hartwig

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Aluminium has been used in a wide range of industrial applications due to its numerous advantages, including excellent specific strength, thermal conductivity, corrosion resistance, workability and recyclability. The automotive industry is increasingly adopting multi-materials, including aluminium in structures and components to improve the mechanical usability and performance of individual components. A common method for assembling dissimilar materials is mechanical joining, but mechanical joining requires multiple manufacturing steps, affects the mechanical properties of the base material and increases the weight due to additional metal parts. Fusion bonding is being used in more and more industries as a way of avoiding the above drawbacks. Infusion bonding, and surface pre-treatment of the base material is essential to ensure the long-life durability of the joint. Laser surface treatment of aluminium has been shown to improve the durability of the joint by forming a passive oxide film and roughening the substrate surface. Infusion bonding, the polymer bonds directly to the metal instead of the adhesive, but the sensitivity to interfacial contamination is higher due to the chemical activity and molecular size of the polymer. Laser-treated surfaces are expected to absorb impurities from the storage atmosphere over time, but the effect of such changes in the treated surface over time on the durability of fusion-bonded joints has not yet been fully investigated. In this paper, the effect of the ageing of laser-treated surfaces of aluminum alloys on the corrosion resistance of fusion-bonded joints is therefore investigated. AlMg3 of 1.5 mm thickness was cut using a water-jet cutting machine, cleaned and degreased with isopropanol and surface pre-treated with a pulsed fiber laser at a wavelength of 1060 nm, maximum power of 70 W and repetition rate of 55 kHz. The aluminum surfaces were then stored in air for various periods of time and their corrosion resistance was assessed by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). For the aluminum joints, induction heating was employed as the fusion bonding method and single-lap shear specimens were prepared. The corrosion resistance of the joints was assessed by measuring the lap shear strength before and after neutral salt spray. Cross-sectional observations by scanning electron microscopy (SEM) were also carried out to investigate changes in the microstructure of the bonded interface. Finally, the corrosion resistance of the surface and the joint were compared and the differences in the mechanisms of corrosion resistance enhancement between the two were discussed.

Keywords: laser surface treatment, pre-treatment, bonding, corrosion, durability, interface, automotive, aluminium alloys, joint, fusion bonding

Procedia PDF Downloads 50

Authors: Rabia Göçmen, Gülşah Kanbur, Sinan Sefa Parlat

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Aim of this study is to determine the effects of cellular insulin receptor stimulators on performance, plasma glucose, high density lipoprotein (HDL), low density lipoprotein (LDL), total cholesterol, triglyceride, triiodothyronine (T3) and thyroxine (T4) hormone levels, and incubation features in the breeding Japanese quails (Coturnix japonica). In the study, a total of 84 breeding quails was used, 6 weeks’ age, 24 are male and 60, female. Rations used in experiment are 2900 kcal/kg metabolic energy and 20% crude protein. Water pH is calibrated to 7.45. Ration and water were administered ad-libitum to the animals. As metformin source, metformin-HCl was used and as chrome resource, chromium picolinate was used. Trial groups were formed as control group (basal ration), metformin group (basal ration, added metformin at the level of feed of 20 mg/kg), and chromium picolinate (basal ration, added feed of 1500 ppb Cr) group. When regarded to the results of performance at the end of experiment, it is seen that live weight gain, feed consumption, egg weight, feed conversion ratio (Feed consumption/ egg weight), and egg production were affected at the significant level (p < 0.05). When the results are evaluated in terms of incubation features, hatchability and hatchability of fertile egg ratio were not affected from the treatments. Fertility ratio was significantly affected by metformin and chromium picolinate treatments and fertility rose at the significant level compared to control group (p < 0.05). According to results of experiment, plasma glucose level was not affected by metformin and chromium picolinate treatments. Plasma, total cholesterol, HDL, LDL, and triglyceride levels were significantly affected from insulin receptor stimulators added to ration (p < 0.05). Hormone level of Plasma T3 and T4 were also affected at the significant level from insulin receptor stimulators added to ration (p < 0.05).

Keywords: chromium picolinate, cholesterol, hormone, metformin, quail

Procedia PDF Downloads 198

Authors: Y. Saylan, F. Yılmaz, A. Denizli

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Rheumatoid arthritis (RA) which is the most common autoimmune disorder of the body's own immune system attacking healthy cells. RA has both articular and systemic effects.Until now romatiod factor (RF) assay is used the most commonly diagnosed RA but it is not specific. Anti-cyclic citrullinated peptide (anti-CCP) antibodies are IgG autoantibodies which recognize citrullinated peptides and offer improved specificity in early diagnosis of RA compared to RF. Anti-CCP antibodies have specificity for the diagnosis of RA from 91 to 98% and the sensitivity rate of 41-68%. Molecularly imprinted polymers (MIP) are materials that are easy to prepare, less expensive, stable have a talent for molecular recognition and also can be manufactured in large quantities with good reproducibility. Molecular recognition-based adsorption techniques have received much attention in several fields because of their high selectivity for target molecules. Quartz crystal microbalance (QCM) is an effective, simple, inexpensive approach mass changes that can be converted into an electrical signal. The applications for specific determination of chemical substances or biomolecules, crystal electrodes, cover by the thin films for bind or adsorption of molecules. In this study, we have focused our attention on combining of molecular imprinting into nanofilms and QCM nanosensor approaches and producing QCM nanosensor for anti-CCP, chosen as a model protein, using anti-CCP imprinted nanofilms. For this aim, anti-CCP imprinted QCM nanosensor was characterized by Fourier transform infrared spectroscopy, atomic force microscopy, contact angle measurements and ellipsometry. The non-imprinted nanosensor was also prepared to evaluate the selectivity of the imprinted nanosensor. Anti-CCP imprinted QCM nanosensor was tested for real-time detection of anti-CCP from aqueous solution. The kinetic and affinity studies were determined by using anti-CCP solutions with different concentrations. The responses related with mass shifts (Δm) and frequency shifts (Δf) were used to evaluate adsorption properties and to calculate binding (Ka) and dissociation (Kd) constants. To show the selectivity of the anti-CCP imprinted QCM nanosensor, competitive adsorption of anti-CCP and IgM was investigated.The results indicate that anti-CCP imprinted QCM nanosensor has a higher adsorption capabilities for anti-CCP than for IgM, due to selective cavities in the polymer structure.

Keywords: anti-CCP, molecular imprinting, nanosensor, rheumatoid arthritis, QCM

Procedia PDF Downloads 341

Authors: Junichi Hosoi

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Epidermal Langerhans cells reside in upper layer of epidermis and play a role in immune surveillance. The finding of the close association of nerve endings to Langerhans cells triggered the research on systemic regulation of Langerhans cells. They disappear from epidermis after exposure to environmental and internal stimuli and reappear about a week later. Myeloid progenitor cells are assumed to be one of the sources of Langerhans cells. We examined the effects of cortisol on the reappearance of Langerhans cells in vitro. Cord-blood derived CD34-positive cells were cultured in the medium supplemented with stem cell factor/Flt3 ligand/granulocyte macrophage-colony stimulating factor/tumor necrosis factor alpha/bone morphologic protein 7/transforming growth factor beta in the presence or absence of cortisol. Cells were analyzed by flow cytometry for CD1a (cluster differentiation 1a), a marker of Langerhans cells and dermal dendritic cells, and CD39 (cluster differentiation factor 39), extracellular adenosine triphosphatase. Both CD1a-positive cells and CD39-positive cells were decreased by treatment with cortisol (suppression by 35% and 22% compared to no stress hormone, respectively). Differentiated Langerhans cells are attracted to epidermis by chemokines that are secreted from keratinocytes. Epidermal keratinocytes were cultured in the presence or absence of cortisol and analyzed for the expression of CCL2 (C-C motif chemokine ligand 2) and CCL20 (C-C motif chemokine ligand 20), which are typical attractants of Langerhans cells, by quantitative reverse transcriptase polymerase chain reaction. The expression of both chemokines, CCL2 and CCL20, were suppressed by treatment with cortisol (suppression by 38% and 48% compared to no stress hormone, respectively). We examined the possible regulation of the suppression by cortisol with plant extracts. The extracts of Ganoderma lucidum and Iris protected the suppression of the differentiation to CD39-positive cells and also the suppression of the gene expression of LC-chemoattractants. These results suggest that cortisol, which is either systemic or locally produced, blocks the supply of epidermal Langerhans cells at 2 steps, differentiation from the precursor and attraction to epidermis. The suppression is possibly blocked by some plant extracts.

Keywords: Langerhans cell, stress, CD39, chemokine

Procedia PDF Downloads 159

Authors: Oubelkacem Yacine, El Bast Hassan, El Bakkali Abdelmajid, Lamhasni Taibi, Ettakni Mahmoud, Ait Lyazidi Saadia, Haddad Mustapha, Ben-Ncer Abdelouahed, El Ferrane Mohammed, Boufarra Abdelkrim

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The present work relates to an on-site and completely non-invasive investigation of one of the most famous west Mediterranean Torah Scroll housed at the National Library of the Kingdom of Morocco. The scroll is 26 m long and consists of 143 parchment sheets of 59 cm x 19 cm, exhibiting only black writings; it is of unknown age. The artifact has been restored by the curator staff of the library. The investigation exploring separately the parchment support and the writing black ink aims at: i) the examination of the parchment conservation/degradation state, ii) the identification of the black ink and iii) the identification of the parchment handcrafting materials. For this purpose, the analyses have been based on combining all of elemental XRF and structural Raman, ATR-FT Infrared Red and Fiber Optical Reflectance spectroscopies, in addition to chroma-metric and pH measurements. pH measurements showing values around 6.5 are in concordance with the absence of any visual corrosion related to the parchment acidity. However, on the basis of the relative intensities and frequency shift of amid I (AI) and amid II (AII) vibrational bands of the collagen, ATR-FTIR spectra revealed diffuse hydrolysis and gelatinization of the parchment writing support; diffuse and non-homogeny degradation by gelatinization has been also confirmed by the IG gelatinization index deduced from the NIR bands on the FOR spectra. This IG index, defined as the ratio I (6860 cm-1) / I (6685 cm-1), ranges in the interval 0.98 – 1 and highlights collagen degradation at the molecular level. Sequentially Shifted Excitation Raman measurements (SSERS) crossed to X-ray fluorescence (XRF) ones on the black writings revealed that the black ink used is an iron-copper gall one, while FOR spectra are typical of pure metal gall inks. These later reflectance measurements exclude, thus, any intentional addition of carbon black to the ink recipe. Moreover, no lead white had been used while pre-drawing the writing lines. On another side, ATR-FTIR measurements highlighted the presence of oxalates as ink degradation products. Considering the parchment handcrafting, the combination of XRF and ATR-FTIR measurements led to the assumption that this writing support had been prepared according to ancient Middle East practices; the parchment infrared fingerprint seems identical to that of the Dead Sea scroll. The present multi-technical analyses are the first ones performed on an ancient Judaic written parchment of Morocco; it is under furthering. The investigation will be extended to other parchments belonging to the Jewish Cultural Heritage Museum of Morocco in Casablanca.

Keywords: torah scroll, parchment, black ink, non-invasive analyses, XRF/ATR-FTIR/RAMAN/FORS

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Authors: Farahnaz Sadat Golestan Hashemi, Mohd Razi Ismail, Mohd Y. Rafii

Abstract:

A huge revolution has emerged in genome engineering by the discovery of CRISPR (clustered regularly interspaced palindromic repeats) and CRISPR-associated system genes (Cas) in bacteria. The function of type II Streptococcus pyogenes (Sp) CRISPR/Cas9 system has been confirmed in various species. Other S. thermophilus (St) CRISPR-Cas systems, CRISPR1-Cas and CRISPR3-Cas, have been also reported for preventing phage infection. The CRISPR1-Cas system interferes by cleaving foreign dsDNA entering the cell in a length-specific and orientation-dependant manner. The S. thermophilus CRISPR3-Cas system also acts by cleaving phage dsDNA genomes at the same specific position inside the targeted protospacer as observed in the CRISPR1-Cas system. It is worth mentioning, for the effective DNA cleavage activity, RNA-guided Cas9 orthologs require their own specific PAM (protospacer adjacent motif) sequences. Activity levels are based on the sequence of the protospacer and specific combinations of favorable PAM bases. Therefore, based on the specific length and sequence of PAM followed by a constant length of target site for the three orthogonals of Cas9 protein, a well-organized procedure will be required for high-throughput and accurate mining of possible target sites in a large genomic dataset. Consequently, we created a reliable procedure to explore potential gRNA sequences for type I (Streptococcus thermophiles), II (Streptococcus pyogenes), and III (Streptococcus thermophiles) CRISPR/Cas systems. To mine CRISPR target sites, four different searching modes of sgRNA binding to target DNA strand were applied. These searching modes are as follows: i) coding strand searching, ii) anti-coding strand searching, iii) both strand searching, and iv) paired-gRNA searching. The output of such procedure highlights the power of comparative genome mining for different CRISPR/Cas systems. This could yield a repertoire of Cas9 variants with expanded capabilities of gRNA design, and will pave the way for further advance genome and epigenome engineering.

Keywords: CRISPR/Cas systems, gRNA mining, Streptococcus pyogenes, Streptococcus thermophiles

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