Search results for: leishmania activated protein kinase c (lack)

Authors: Sonu Gandhi, Neena Capalash, Prince Sharma, C. Raman Suri

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A sensitive chemiluminescence immunoassay (CIA) for heroin and its major metabolites is reported. The method is based on the competitive reaction of horseradish peroxidase (HRP)-labeled anti-MAM antibody and free drug in spiked urine samples. A hapten-protein conjugate was synthesized by using acidic derivative of monoacetyl morphine (MAM) coupled to carrier protein BSA and was used as an immunogen for the generation of anti-MAM (monoacetyl morphine) antibody. A high titer of antibody (1:64,0000) was obtained and the relative affinity constant (Kaff) of antibody was 3.1×107 l/mol. Under the optimal conditions, linear range and reactivity for heroin, mono acetyl morphine (MAM), morphine and codeine were 0.08, 0.09, 0.095 and 0.092 ng/mL respectively. The developed chemiluminescence inhibition assay could detect heroin and its metabolites in standard and urine samples up to 0.01 ng/ml.

Keywords: heroin, metabolites, chemiluminescence immunoassay, horse radish peroxidase

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Authors: Iulianna Taritsa, Kuldeep Neote, Eric Fossel

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Lysosome-induced immunogenic cell death (LIICD) is a powerful mechanism of targeting cancer cells that kills circulating malignant cells and primes the host’s immune cells against future remission. Current immunotherapies for cancer are limited in preventing recurrence – a gap that can be bridged by training the immune system to recognize cancer neoantigens. Lysosomal leakage can be induced therapeutically to traffic antigens from dying cells to dendritic cells, which can later present those tumorigenic antigens to T cells. Previous research has shown that oxidative agents administered in the tumor microenvironment can initiate LIICD. We generated a fusion protein between an oxidative agent known as xanthine oxidase (XO) and a mini-antibody specific for EGFR/HER2-sensitive breast tumor cells. The anti-EGFR single domain antibody fragment is uniquely sourced from llama, which is functional without the presence of a light chain. These llama micro-antibodies have been shown to be better able to penetrate tissues and have improved physicochemical stability as compared to traditional monoclonal antibodies. We demonstrate that the fusion protein created is stable and can induce early markers of immunogenic cell death in an in vitro human breast cancer cell line (SkBr3). Specifically, we measured overall cell death, as well as surface-expressed calreticulin, extracellular ATP release, and HMGB1 production. These markers are consensus indicators of ICD. Flow cytometry, luminescence assays, and ELISA were used respectively to quantify biomarker levels between treated versus untreated cells. We also included a positive control group of SkBr3 cells dosed with doxorubicin (a known inducer of LIICD) and a negative control dosed with cisplatin (a known inducer of cell death, but not of the immunogenic variety). We looked at each marker at various time points after cancer cells were treated with the XO/antibody fusion protein, doxorubicin, and cisplatin. Upregulated biomarkers after treatment with the fusion protein indicate an immunogenic response. We thus show the potential for this fusion protein to induce an anticancer effect paired with an adaptive immune response against EGFR/HER2+ cells. Our research in human cell lines here provides evidence for the success of the same therapeutic method for patients and serves as the gateway to developing a new treatment approach against breast cancer.

Keywords: apoptosis, breast cancer, immunogenic cell death, lysosome

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Authors: Anshuman Srivastava, Shiv Singh, Sheelendra Pratap Singh

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A simple, sensitive and effective gas chromatography tandem mass spectrometry (GC-MS/MS) method was developed for simultaneous analysis of multi pesticide residues (organophosphate, organochlorines, synthetic pyrethroids and herbicides) in food commodities using phenolic resin based activated carbon fibers (ACFs) as reversed-dispersive solid phase extraction (r-DSPE) sorbent in modified QuEChERS (Quick Easy Cheap Effective Rugged Safe) method. The acetonitrile-based QuEChERS technique was used for the extraction of the analytes from food matrices followed by sample cleanup with ACFs instead of traditionally used primary secondary amine (PSA). Different physico-chemical characterization techniques such as Fourier transform infrared spectroscopy, scanning electron microscopy, X-ray diffraction and Brunauer-Emmet-Teller surface area analysis were employed to investigate the engineering and structural properties of ACFs. The recovery of pesticides and herbicides was tested at concentration levels of 0.02 and 0.2 mg/kg in different commodities such as cauliflower, cucumber, banana, apple, wheat and black gram. The recoveries of all twenty-six pesticides and herbicides were found in acceptable limit (70-120%) according to SANCO guideline with relative standard deviation value < 15%. The limit of detection and limit of quantification of the method was in the range of 0.38-3.69 ng/mL and 1.26 -12.19 ng/mL, respectively. In traditional QuEChERS method, PSA used as r-DSPE sorbent plays a vital role in sample clean-up process and demonstrates good recoveries for multiclass pesticides. This study reports that ACFs are better in terms of removal of co-extractives in comparison of PSA without compromising the recoveries of multi pesticides from food matrices. Further, ACF replaces the need of charcoal in addition to the PSA from traditional QuEChERS method which is used to remove pigments. The developed method will be cost effective because the ACFs are significantly cheaper than the PSA. So the proposed modified QuEChERS method is more robust, effective and has better sample cleanup efficiency for multiclass multi pesticide residues analysis in different food matrices such as vegetables, grains and fruits.

Keywords: QuEChERS, activated carbon fibers, primary secondary amine, pesticides, sample preparation, carbon nanomaterials

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Authors: Hosam El-Sayed, Amira Abou El-Kheir, Salwa Mowafi, Marwa Abou Taleb

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Two proteinic biopolymers; namely keratin and sericin, were extracted from their respective natural resources by simple appropriate methods. The said proteins were dissolved in the appropriate solvents followed by regeneration in a form of film polyvinyl alcohol. Proteinium thiocyanate (PTC) composite was prepared by reaction of a regenerated film with potassium thiocyanate in acid medium. In another experiment, the said acidified proteins were reacted with potassium thiocyante before dissolution and regeneration in a form of PTC composite. The possibility of using PTC composite for determination of the concentration of iron III ions in domestic as well as industrial water was examined. The concentration of iron III cations in water was determined spectrophotometrically by measuring the intensity of blood red colour of iron III thiocyanate obtained by interaction of PTC with iron III cation in the tested water sample.

Keywords: iron III cations, protein, sensor, thiocyanate, water

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Authors: Seemab Zehra, A. H. W. Mohammed, E. Pantanella, J. L. Q. Laranja, P. H. De Mello, R. Saleh, A. A. Siddik, A. Al Shaikhi, A. M. Al-Suwailem

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Two separate feeding trials were conducted to determine the effects of using different commercial diets and water temperatures on the growth performance, feed intake, feed conversion ratio (FCR) and condition factor of sobaity seabream Sparidentex hasta. In experiment I, growth performance, feed intake, protein efficiency ratio (PER), feed conversion ratio (FCR) and survival (%) of sobaity seabream Sparidentex hasta (330.5±2.6 g; 26.9±1.0 cm) were evaluated by four different commercial diets (1, 2, 3 and 4) for 80 days. The daily weight gain was around 3.2 g day-1 with an SGR of 0.7% day-1. Both the FCR and PER in the fish were significantly better in diet 2 that contained 46.36% crude protein and 12.54% crude fat. In experiment II, (99±2.6 g; 17.1±1.0 cm). The fish were cultured in 1m3 tanks supplied with seawater from the Red Sea wherein three different rearing temperatures were set as treatments (24, 28 and 32°C). Fish were fed with a commercial diet based on the results of experiment I (46.4% protein; 20.1 MJ kg-1 energy) to satiation for 96 days. Total weight gain was significantly higher for the fish reared in the 32°C group (158.57 g) followed by the 28°C group (138.25 g), while the lowest weight gain was observed in the 24°C group (116.98 g). The FCR was significantly lower in the 32°C group (1.62) as compared to 28 (1.8) and 24°C (1.85) groups. Based on the results obtained from these preliminary studies (experiment I and II), sobaity seabream can attain better growth performance, FCR and PER at 32°C in the Red Sea by feeding commercial diet 2.

Keywords: Sparidentex hasta, nutrition, FCR, Red Sea, growth performance

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Authors: Zahid Manzoor, Shaukat Hussain Munawar, Zahid Iqbal, Imran Ahmad Khan, Abdul Aziz, Hafiz Muhammad Qasim

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The present study was aimed to investigate the pharmacokinetics behavior and optimal dosage regimen of danofloxacin in 8 adult healthy buffaloes of local breed (Nili Ravi) following single intravenous administration at the dose of 2.5 mg/kg body weight. Plasma drug concentrations at various time intervals were measured by HPLC method. In vitro plasma protein binding was determined employing the ultrafiltration technique. The distribution and elimination of danofloxacin was rapid, as indicated by the values (Mean±SD) of distribution half-life (t1/2α = 0.25±0.09 hours) and elimination half life (t1/2β = 3.26±0.43 hours), respectively. Volume of distribution at steady state (Vss) was 1.14±0.12 L/kg, displaying its extensive distribution into various body fluids and tissues. The high value of AUC (9.80±2.14 µg/ml.hr) reflected the vast area of the body covered by drug concentration. The mean residence time was noted to be 4.78±0.52 hours. On the basis of pharmacokinetic parameters, a suitable intravenous regimen for danofloxacin in adult buffaloes would be 6.5 mg/kg to be repeated after 12 hours intervals. The present study is the foremost pharmacokinetic study of danofloxacin in the local species which would provide the valueable contribution in the local manufacturing of danofloxacin in Pakistan in future.

Keywords: danofloxacin, pharmacokinetics, plasma protein binding, buffaloes, dosage regimen

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Authors: Masoud Rafiee

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To study the quality response of forage to chickpea-barley mixed cropping under drought stress and vermicompost consumption, an experiment was carried out under well watered and %70 water requirement (stress condition) in RCBD as split plot with four replications in temperate condition of Khorramabad in 2013. Chickpea-barley mix cropping (%100 chickpea, %75:25 chickpea:barley, %50:50 chickpea:barley, %25:75 chickpea:barley, and %100 barley) was studied. Results showed that wet and dry forage yield were significantly affected by environment and decreased in stress condition. Also, crude protein content decreased from %26.2 in well watered to %17.3 in stress condition.

Keywords: crude protein, wet forage yield, dry forage yield, water stress condition, well watered

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Authors: Makhan Maharjan, Mani Ulaganathan, Vanchiappan Aravindan, Srinivasan Madhavi, Jing-Yuan Wang, Tuti Mariana Lim

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There is great interest to exploit sustainable, low-cost, renewable resources as carbon precursors for energy storage applications. Research on development of energy storage devices has been growing rapidly due to mismatch in power supply and demand from renewable energy sources This paper reported the synthesis of porous activated carbon from biomass waste and evaluated its performance in supercapicators. In this work, we employed orange peel (waste material) as the starting material and synthesized activated carbon by pyrolysis of KOH impregnated orange peel char at 800 °C in argon atmosphere. The resultant orange peel-derived activated carbon (OP-AC) exhibited a high BET surface area of 1,901 m2 g-1, which is the highest surface area so far reported for the orange peel. The pore size distribution (PSD) curve exhibits the pores centered at 11.26 Å pore width, suggesting dominant microporosity. The OP-AC was studied as positive electrode in combination with different negative electrode materials, such as pre-lithiated graphite (LiC6) and Li4Ti5O12 for making different hybrid capacitors. The lithium ion capacitor (LIC) fabricated using OP-AC with pre-lithiated graphite delivered a high energy density of ~106 Wh kg–1. The energy density for OP-AC||Li4Ti5O12 capacitor was ~35 Wh kg–1. For comparison purpose, configuration of OP-AC||OP-AC capacitors were studied in both aqueous (1M H2SO4) and organic (1M LiPF6 in EC-DMC) electrolytes, which delivered the energy density of 6.6 Wh kg-1 and 16.3 Wh kg-1, respectively. The cycling retentions obtained at current density of 1 A g–1 were ~85.8, ~87.0 ~82.2 and ~58.8% after 2500 cycles for OP-AC||OP-AC (aqueous), OP-AC||OP-AC (organic), OP-AC||Li4Ti5O12 and OP-AC||LiC6 configurations, respectively. In addition, characterization studies were performed by elemental and proximate composition, thermogravimetry, field emission-scanning electron microscopy, Raman spectra, X-ray diffraction (XRD) pattern, Fourier transform-infrared, X-ray photoelectron spectroscopy (XPS) and N2 sorption isotherms. The morphological features from FE-SEM exhibited well-developed porous structures. Two typical broad peaks observed in the XRD framework of the synthesized carbon implies amorphous graphitic structure. The ratio of 0.86 for ID/IG in Raman spectra infers high degree of graphitization in the sample. The band spectra of C 1s in XPS display the well resolved peaks related to carbon atoms in various chemical environments; for instances, the characteristics binding energies appeared at ~283.83, ~284.83, ~286.13, ~288.56, and ~290.70 eV which correspond to sp2 -graphitic C, sp3 -graphitic C, C-O, C=O and π-π*, respectively. Characterization studies revealed the synthesized carbon to be promising electrode material towards the application for energy storage devices. The findings opened up the possibility of developing high energy LICs from abundant, low-cost, renewable biomass waste.

Keywords: lithium-ion capacitors, orange peel, pre-lithiated graphite, supercapacitors

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Authors: Emely J. Escala, Lolito C. Bestil

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An experiment was conducted to assess the potential of jackfruit by-product concentrate (JBC) in combination with two types of protein sources, soybean meal (SBM) and liquid acid whey (LAW), given at two different ratios as supplement for growing kids fed a basal diet of 70:30 napier grass (Pennisetum purpureum) and kakawate (Gliricidia sepium) soilage ratio. The experiment was set-up in randomized complete block design (RCBD) with sex-age combination as basis for blocking, with the following dietary treatments: T1 = 0.50:0.50% BW, DM basis, JBC:SBM, T2 = 0.75:0.25% BW JBC:SBM, T3 = 0.50:0.50% BW, DM basis, JBC:LAW, and T4 = 0.75:0.25% BW JBC:LAW. Analysis of JBC showed high contents of crude fiber with medium levels of crude protein and nitrogen-free extract, appearing to be fitting for ruminants and a potential energy source. Results showed significantly higher voluntary dry matter intake (VDMI), cumulative weight gain (CWG), and average daily gain (ADG) of growing goats supplemented with JBC in combination with SBM than with LAW. The amount of JBC can range from 0.50% to 0.75% BW with SBM making up the difference, but a JBC:SBM ratio of 0.75:0.25% BW, DM basis, is best in promoting highest voluntary dry matter intake and is, therefore, highly recommended in the light of savings in feed cost. A long-term study on the effects of JBC supplementation on meat qualities of growing kids (aroma, marbling characteristics and taste) is also recommended.

Keywords: jackfruit by-product concentrate, liquid acid whey, soybean meal, grower kids

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Authors: Sara Taghdisi, M. Mirmohammadi, M. Mokhtarian

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In 2013, the World Health Organization announced the cases of Cutaneous leishmaniasis infection in Iran between 69,000 to 113,000. The most common chemical drugs for Cutaneous leishmaniasis treatment are sodium stibogluconate, and meglumine antimonate, which not only have relatively many side effects, but also some species of the Leishmania genus have become resistant to them .The most prominent compound existing in different parts of the cotton plant is a yellow polyphenol called Gossypol. Gossypol is an extremely valuable compound and has anti-cancer properties. In the current project, Gossypol was extracted with a liquid-liquid extraction method in 120 minutes in the presence of Phosphoric acid from the cotton seed oil of Golestan beach varieties, then got crystallized in darkness using Acetic acid and isolated as Gossypol Acetic acid. The efficiency of the extracted crystal was obtained at 0.12+- 1.28. the cotton plant could be efficient in the treatment of Cutaneous leishmaniasis. The extract of the green-leaf cotton boll of Jargoyeh varieties was tested as an ointment on the target group of patients suffering from Cutaneous leishmaniasis resistant to drugs esistant to drugs by our colleagues in the research team. The results showed the Pearson's correlation coefficient of 0.72 between the two variables of wound diameter and the extract use over time which indicated the positive effect of this extract on the treatment of Cutaneous leishmaniasis was resistant to drugs.

Keywords: cottonseed oil, crystallization, gossypol, green-leaf

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Authors: Shivangi Agrawal, Hsiu-I Ting

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Entrepreneurship is an essential source of economic growth and a prominent factor influencing socio-economic development. Entrepreneurship education educates and enhances entrepreneurial activity. This study aims to understand current trends in entrepreneurship education and evaluate the effectiveness of diverse entrepreneurship education programs. An increasing number of universities offer entrepreneurship education courses to create and successfully continue entrepreneurial ventures. Despite the prevalence of entrepreneurship education, research studies lack inconsistency about the effectiveness of entrepreneurship education to promote and develop entrepreneurship. Strategies to develop entrepreneurial attitudes and intentions among individuals are hindered by a lack of understanding of entrepreneurs' educational purposes, components, methodology, and resources required. Lack of adequate entrepreneurship education has been linked with low self-efficacy and lack of entrepreneurial intent. Moreover, in the age of digitisation and during the COVID-19 pandemic, digital learning platforms (e.g., online entrepreneurship education courses and programs) and other digital tools (e.g., digital game-based entrepreneurship education) have become more relevant to entrepreneurship education. This paper contributes to the continuation of academic literature in entrepreneurship education by evaluating and assessing current trends in entrepreneurship education programs, leading to better understanding to reduce gaps between entrepreneurial development requirements and higher education institutions.

Keywords: entrepreneurship education, digital technologies, academic entrepreneurship, COVID-19

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Authors: Shahnaz Mansouri, Xiao Dong Chen, Meng Wai Woo

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A new antisolvent vapour precipitation approach was used to make ultrafine submicron probiotic encapsulates. The approach uses ethanol vapour to precipitate submicron encapsulates within relatively large droplets. Surprisingly, the probiotics (Lactobacillus delbrueckii ssp. bulgaricus, Streptococcus thermophilus) showed relatively high survival even under destructive ethanolic conditions within the droplet. This unusual behaviour was deduced to be caused by the denaturation and aggregation of the milk protein forming an ethanolic protective matrix for the probiotics. Skim milk droplets which is rich in casein and contains naturally occurring minerals provided higher ethanolic protection when compared whey protein isolate and lactose droplets.

Keywords: whey, skim milk, probiotic, antisolvent, precipitation, encapsulation, denaturation, aggregation

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Authors: Homa Torabizadeh, Mohaddeseh Mikani

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Inulinase from Aspergillus niger was covalently immobilized on magnetic nanoparticles (MNPs/Fe₃O₄) covered with soy protein isolate (SPI/Fe₃O₄) functionalized by bovine serum albumin (BSA) nanoparticles. MNPs are promising enzyme carriers because they separate easily under external magnetic fields and have enhanced immobilized enzyme reusability. As MNPs aggregate simply, surface coating strategy was employed. SPI functionalized by BSA was a suitable candidate for nanomagnetite coating due to its superior biocompatibility and hydrophilicity. Fe₃O₄@SPI-BSA nanoparticles were synthesized as a novel carrier with narrow particle size distribution. Step by step fabrication monitoring of Fe₃O₄@SPI-BSA nanoparticles was performed using field emission scanning electron microscopy and dynamic light scattering. The results illustrated that nanomagnetite with the spherical morphology was well monodispersed with the diameter of about 35 nm. The average size of the SPI-BSA nanoparticles was 80 to 90 nm, and their zeta potential was around −34 mV. Finally, the mean diameter of fabricated Fe₃O₄@SPI-BSA NPs was less than 120 nm. Inulinase enzyme from Aspergillus niger was covalently immobilized through gluteraldehyde on Fe₃O₄@SPI-BSA nanoparticles successfully. Fourier transform infrared spectra and field emission scanning electron microscopy images provided sufficient proof for the enzyme immobilization on the nanoparticles with 80% enzyme loading.

Keywords: high fructose syrup, inulinase immobilization, functionalized magnetic nanoparticles, soy protein isolate

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

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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

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Authors: Kavita Jaidiya, Anju Chahar, Chitra Jaidiya

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The present study was conducted on 200 quarters from 50 apparently healthy cows. Samples are subjected to California Mastitis Test (CMT), cultural examination, and mPCR. Milk samples were also subjected to changes in composition Viz. fat, protein, and lactose. The prevalence of subclinical mastitis based on culture examination was 30(60/200), 36 (72/200), and 40 percent (93/200) based on CMT, culture examination, and mPCR on a quarterly basis. The prevalence of subclinical mastitis on animal basis was 40 (20/50), 46 (23/50), and 52 percent (26/50) based on CMT, Culture examination, and mPCR. The highest prevalence was observed in IVth parity on a quarterly basis and in Vth parity on cow basis. On culture examination, Staphylococcus aureus was the most prevalent organism (50.56%), followed by Streptococcus dysaglactiae (11.33%), E. coli (7.8 %), Staphylococcus agalactiae (13.48 %), Staphylococcus epidermidis (2.2 %), Streptococcus hyicus (6.94%), Streptococcus uberis (5.16%), Klebsiella pneumonia (6.74%). On isolation by bacterial mPCR, Staphylococcus spp. (42%) was the major pathogen. Organisms isolated in mixed infections are Streptococcus spp., Klebsiella pneumonia, E.coli and Pseudomonas aeruginous. The average mean value of fat, protein, and lactose content in subclinically affected milk samples were 3.40 ± 0.101, 3.009 ± 0.033, and 4.48 ± 0.03, and the mean value of fat, protein, and lactose content in normal milk were 4.13 ± 0.035, 3.39 ± 0.021, and 5.10 ± 0.016. The mean blood level of reduced glutathione in subclinical mastitis (30.44 ± 1.87 ng/ml) was lower than healthy cows (47.98 ± 4.04ng/ml). The concentration of malondialdehyde (10.026 ± 0.21mmol/L) in subclinical mastitis was significantly higher as compared to healthy group cows (2.19 ± 0.23mmol/L).

Keywords: cow, subclinical mastitis, mPCR, California Mastitis test

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Authors: Christopher Wearmouth, Jacob Smith

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Administration of tranexamic acid (TXA) reduces all-cause mortality in trauma patients when given within 3 hours of injury. Due to geographical distance and lack of emergency medical services patients often present late, following trauma, to our emergency department. Additionally, we found patients that may have benefited from TXA did not receive it, often due to lack of staff awareness, staff shortages out of hours and lack of equipment for delivering infusions. Our objective was to develop a protocol for nurse-led administration of TXA in the emergency department. We developed a protocol using physiological observations along with criteria from the South African Triage Scale to allow nursing staff to identify patients with, or at risk of, significant haemorrhage. We will monitor the use of the protocol to ensure appropriate compliance and for any adverse events reported.

Keywords: emergency department, emergency nursing, rural healthcare, tranexamic acid, trauma, triage

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Authors: Chanya Inprasit, Yi-Wen Lin

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Inflammation causes changes of peripheral and central nervous system properties, affecting both neuronal and non-neuronal cells, resulting in inflammatory pain. Acupoint injection (AI) was developed in the 1950s and has been widely used for relieving pain. It is an acupoint-stimulating technique that utilizes anatomically based meridians derived from Chinese medicine theory. AI has been accepted as an effective treatment and is thought to display superior results when compared to traditional acupuncture methods. However, the mechanism of AI needs to be ratified by more scientific evidence in order to support the theory and its therapeutic development. In this study, we explored the effect of AI on the comorbidity of chronic pain and depression. Mice hindpaw was injected by complete Freund’s adjuvant (CFA) to induce the condition of chronic pain. Measurements of mechanical and thermal hyperalgesia and depression-like behavior were analyzed. The results indicated a positive tendency to AI treatment. The comorbidity of chronic pain and depression was investigated with relation to transient receptor potential V1 (TRPV1) mechanism through the use of TRPV1 gene deletion. The expression of nociceptors such as voltage-gated sodium channels (Navs) or TRPV1, was significantly down-regulated by AI. The expression of inflammation-activated molecules: astrocytic marker glial fibrillary acidic protein (GFAP), the microglial marker Iba-1, S100B, and related kinases, were reversed by AI in both the peripheral and central nervous system. Taken together, these data provided a detailed molecular mechanism of AI-induced analgesia and anti-inflammatory properties. This finding may be utilized for clinical practice to treat chronic pain and depression comorbidity.

Keywords: inflammatory pain, acupoint injection, TRPV1, GFAP, S100B

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Authors: Chaoqun Tan, Naiyun Gao, Xiaoyan Xin

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Perxymonosulfate (PMS)-based oxidation processes, as an alternative of hydrogen peroxide-based oxidation processes, are more and more popular because of reactive radical species (SO4-•, OH•) produced in systems. Magnetic nano-scaled particles Fe3O4 and ferrous anion (Fe2+) were studied for the activation of PMS for degradation of acetaminophen (APAP) in water. The Fe3O4 MNPs were found to effectively catalyze PMS for APAP and the reactions well followed a pseudo-first-order kinetics pattern (R2>0.95). While the degradation of APAP in PMS-Fe2+ system proceeds through two stages: a fast stage and a much slower stage. Within 5 min, approximately 7% and 18% of 10 ppm APAP was accomplished by 0.2 mM PMS in Fe3O4 (0.8g/L) and Fe2+ (0.1mM) activation process. However, as reaction proceed to 120 min, approximately 75% and 35% of APAP was removed in Fe3O4 activation process and Fe2+ activation process, respectively. Within 120 min, the mineralization of APAP was about 7.5% and 5.0% (initial APAP of 10 ppm and [PMS]0 of 0.2 mM) in Fe3O4-PMS and Fe2+-PMS system, while the mineralization could be greatly increased to about 31% and 40% as [PMS]0 increased to 2.0 mM in in Fe3O4-PMS and Fe2+-PMS system, respectively. At last, the production of reactive radical species were validated directly from Electron Paramagnetic Resonance (ESR) tests with 0.1 M 5,5-Dimethyl-1-pyrrolidine N-oxide (DMPO). Plausible mechanisms on the radical generation from Fe3O4 and Fe2+ activation of PMS are proposed on the results of radial identification tests. The results demonstrated that Fe3O4 MNPs activated PMS and Fe2+ anion activated PMS systems are promising technologies for water pollution caused by contaminants such as pharmaceutical. Fe3O4-PMS system is more suitable for slowly remediation, while Fe2+-PMS system is more suitable for fast remediation.

Keywords: acetaminophen, peroxymonosulfate, radicals, Electron Paramagnetic Resonance (ESR)

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Authors: Richa Mardianingrum, Srie R. N. Endah

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In this research, we have designed four isoniazid derivatives i.e., isonicotinohydrazide (1-isonicotinoyl semicarbazide, 1-thiosemi isonicotinoyl carbazide, N '-(1,3-dimethyl-1 h-pyrazole-5-carbonyl) isonicotino hydrazide, and N '-(1,2,3- 4-thiadiazole-carbonyl) isonicotinohydrazide. The docking and molecular dynamic have performed to them in order to study its interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (InhA). Based on this research, all of the compounds were predicted to have a stable interaction with Mycobacterium tuberculosis Enoyl-Acyl Carrier Protein Reductase (INHA) receptor, so they could be used as an anti-tuberculosis drug candidate.

Keywords: anti-tuberculosis, docking, Inhibin alpha subunit, InhA, inhibition, synthesis, isonicotinohydrazide

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Authors: Fatima Zohra Bissaad, Farid Bounaceur, Nassima Behidj, Nadjiba Chebouti, Fatma Halouane, Bahia Doumandji-Mitiche

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Effect of Beauveria bassiana and Metarhizium anisopliae var. acridum on the 5^th instar nymphs of Schistocerca gregaria was studied in the laboratory. Infection by these both entomopathogenic fungi caused reduction in the hemolymph total protein. The average amounts of total proteins were 2.3, 2.07, 2.09 µg/100 ml of haemolymph in the control and M. anisopliae var. acridum, and B. bassiana based-treatments, respectively. Three types of haemocytes were recognized and identified as prohaemocytes, plasmatocytes and granulocytes. The treatment caused significant reduction in the total haemocyte count and in each haemocyte type on the 9^th day after its application.

Keywords: Beauveria bassiana, haemolymph picture, haemolymph protein, Metarhizium anisopliae var. acridum, Schistocerca gregaria

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

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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

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Authors: Lee A. Browne, K. Rumbold

Abstract:

The first fluorinating enzyme, 5'-fluoro-5'-deoxyadenosine synthase (fluorinase) was isolated from the soil bacterium Streptomyces cattleya. Such an enzyme, with the ability to catalyze a C-F bond, presents great potential as a biocatalyst. Naturally fluorinated compounds are extremely rare in nature. As a result, the number of fluorinases identified remains relatively few. The field of fluorination is almost completely synthetic. However, with the increasing demand for fluorinated organic compounds of commercial value in the agrochemical, pharmaceutical and materials industries, it has become necessary to utilize biologically based methods such as biocatalysts. A key step in this crucial process is the large-scale production of the fluorinase enzyme in considerable quantities for industrial applications. Thus, this study aimed to optimize expression of the fluorinase enzyme in both prokaryotic and eukaryotic expression systems in order to obtain high protein yields. The fluorinase gene was cloned into the pET 41b(+) and pPinkα-HC vectors and used to transform the expression hosts, E.coli BL21(DE3) and Pichia pastoris (PichiaPink™ strains) respectively. Expression trials were conducted to select optimal conditions for expression in both expression systems. Fluorinase catalyses a reaction between S-adenosyl-L-Methionine (SAM) and fluoride ion to produce 5'-fluorodeoxyadenosine (5'FDA) and L-Methionine. The activity of the enzyme was determined using HPLC by measuring the product of the reaction 5'FDA. A gradient mobile phase of 95:5 v/v 50mM potassium phosphate buffer to a final mobile phase containing 80:20 v/v 50mM potassium phosphate buffer and acetonitrile were used. This resulted in the complete separation of SAM and 5’-FDA which eluted at 1.3 minutes and 3.4 minutes respectively. This proved that the fluorinase enzyme was active. Optimising expression of the fluorinase enzyme was successful in both E.coli and PichiaPink™ where high expression levels in both expression systems were achieved. Protein production will be scaled up in PichiaPink™ using fermentation to achieve large-scale protein production. High level expression of protein is essential in biocatalysis for the availability of enzymes for industrial applications.

Keywords: biocatalyst, expression, fluorinase, PichiaPink™

Procedia PDF Downloads 552

Authors: Soo Dong Cho, In-Ohk Ouh, Byeong Sul Kang, Seyeon Park, In-Soo Cho, Jae Young Song

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An VP2 gene from the current prevalent CPV (Canine Parvovirus) strain (new CPV-2a) in the Republic of Korea was expressed in a baculovirus expression system. Genomic DNA was extracted from the isolate strain CPV-2a. The recombinant baculovirus, containing the coding sequences of VP2 with the histidine tag at the N-terminus, were generated by using the Bac-to-Bac system. For production of the recombinant VP2 proteins, SF9 cells were transfection into 6 wells. Propagation of recombinant baculoviruses and expression of the VP2 protein were performed in the Sf9 cell line maintained. The proteins were detected to Western blot anlaysis. CPV-2a VP2 was detected by Western blotting the monoclonal antibodies recognized 6x His and the band had a molecular weight of 65 KDa. We demonstrated that recombinant CPV-2a VP2 expression in baculovirus. The recombinant CPV-2a VP2 may able to development of specific diagnostic test and vaccination of against CPV2. This study provides a foundation for application of CPV2 on the development of new CPV2 subunit vaccine.

Keywords: baculovirus, canine parvovirus 2a, Dog, Korea

Procedia PDF Downloads 244

Authors: A. M. Nour, M. A. Zaki, E. A. Omer, Nourhan Mohamed

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Performances of mixed-sex Nile tilapia (Oreochromis niloticus) fingerlings (11.33 ± 1.78 g /fish) reared under biofloc system developed by molasses and sucrose as carbon sources in indoor fiberglass tanks were evaluated. Six indoor fiberglass tanks (1m 3 each filled with 1000 l of underground fresh water), each was stocked with 2kg fish were used for 14 weeks experimental period. Three experimental groups were designed (each group 2 tanks) as following: 1-control: 20% daily without biofloc, 2-zero water exchange rate with biofloc (molasses as C source) and 3-zero water exchange rate with biofloc (sucrose as C source). Fish in all aquariums were fed on floating feed pellets (30% crude protein, 3 mm in diameter) at a rate of 3% of the actual live fish body, 3 times daily and 6 days a week. Carbohydrate supplementations were applied daily to each tank two hrs, after feeding to maintain the carbon: nitrogen ratio (C: N) ratio 20:1. Fish were reared under continuous aeration by pumping air into the water in the tank bottom using two sandy diffusers and constant temperature between 27.0-28.0 ºC by using electrical heaters for 10 weeks. Criteria's for assessment of water quality parameters, biofloc production and fish growth performances were collected and evaluated. The results showed that total ammonia nitrogen in control group was higher than biofloc groups. The biofloc volumes were 19.13 mg/l and 13.96 mg/l for sucrose and molasses, respectively. Biofloc protein (%), ether extract (%) and gross energy (kcal/100g DM), they were higher in biofloc molasses group than biofloc sucrose group. Tilapia growth performances were significantly higher (P < 0.05) with molasses group than in sucrose and control groups, respectively. The highest feed and nutrient utilization values for protein efficiency ratio (PER), protein productive (PPV%) and energy utilization (EU, %) were higher in molasses group followed by sucrose group and control group respectively.

Keywords: biofloc, Nile tilapia, cabohydrates, performances

Procedia PDF Downloads 192

Authors: Yuan-Chung Tsai, Masao Kamimura, Kohei Soga, Hsin-Cheng Chiu

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In order to enhance the photodynamic/photothermal therapeutic efficacy on glioblastoma, the functionized upconversion nanoparticles with the capability of converting the deep tissue penetrating near-infrared light into visible wavelength for activating photochemical reaction were developed. The drug-loaded nanoparticles (NPs) were obtained from the self-assembly of oleic acid-coated upconversion nanoparticles along with maleimide-conjugated poly(ethylene glycol)-cholesterol (Mal-PEG-Chol), as the NP stabilizer, and hydrophobic photosensitizers, IR-780 (for photothermal therapy, PTT) and mTHPC (for photodynamic therapy, PDT), in aqueous phase. Both the IR-780 and mTHPC were loaded into the hydrophobic domains within NPs via hydrophobic association. The peptide targeting ligand, angiopep-2, was further conjugated with the maleimide groups at the end of PEG adducts on the NP surfaces, enabling the affinity coupling with the low-density lipoprotein receptor-related protein-1 of tumor endothelial cells and malignant astrocytes. The drug-loaded NPs with the size of ca 80 nm in diameter exhibit a good colloidal stability in physiological conditions. The in vitro data demonstrate the successful targeting delivery of drug-loaded NPs toward the ALTS1C1 cells (murine astrocytoma cells) and the pronounced cytotoxicity elicited by combinational effect of PDT and PTT. The in vivo results show the promising brain orthotopic tumor targeting of drug-loaded NPs and sound efficacy for brain tumor dual-modality treatment. This work shows great potential for improving photodynamic/photothermal therapeutic efficacy of brain cancer.

Keywords: drug delivery, orthotopic brain tumor, photodynamic/photothermal therapies, upconversion nanoparticles

Procedia PDF Downloads 195

Authors: Josef Drahorad, Milos Beran, Ondrej Vltavsky, Marian Urban, Martin Fronek, Jiri Sova

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Engineering efforts of researchers of the Food research institute Prague and the Czech Technical University in spray drying technologies led to the introduction of a demonstrator ATOMIZER and a new technology of Carbon Dioxide-Assisted Spray Nebulization Drying (CASND). The equipment combines the spray drying technology, when the liquid to be dried is atomized by a rotary atomizer, with Carbon Dioxide Assisted Nebulization - Bubble Dryer (CAN-BD) process in an original way. A solution, emulsion or suspension is saturated by carbon dioxide at pressure up to 80 bar before the drying process. The atomization process takes place in two steps. In the first step, primary droplets are produced at the outlet of the rotary atomizer of special construction. In the second step, the primary droplets are divided in secondary droplets by the CO2 expansion from the inside of primary droplets. The secondary droplets, usually in the form of microbubbles, are rapidly dried by warm air stream at temperatures up to 60ºC and solid particles are formed in a drying chamber. Powder particles are separated from the drying air stream in a high efficiency fine powder separator. The product is frequently in the form of submicron hollow spheres. The CASND technology has been used to produce microparticulated protein concentrates for human nutrition from alternative plant sources - hemp and canola seed filtration cakes. Alkali extraction was used to extract the proteins from the filtration cakes. The protein solutions after the alkali extractions were dried with the demonstrator ATOMIZER. Aerosol particle size distribution and concentration in the draying chamber were determined by two different on-line aerosol spectrometers SMPS (Scanning Mobility Particle Sizer) and APS (Aerodynamic Particle Sizer). The protein powders were in form of hollow spheres with average particle diameter about 600 nm. The particles were characterized by the SEM method. The functional properties of the microparticulated protein concentrates were compared with the same protein concentrates dried by the conventional spray drying process. Microparticulated protein has been proven to have improved foaming and emulsifying properties, water and oil absorption capacities and formed long-term stable water dispersions. This work was supported by the research grants TH03010019 of the Technology Agency of the Czech Republic.

Keywords: carbon dioxide-assisted spray nebulization drying, canola seed, hemp seed, microparticulated proteins

Procedia PDF Downloads 169

Authors: Nafees Ahmed, Nur Izyani Kamaruzman, Saralla Nathan, Mohd Ezharul Hoque Chowdhury, Anuar Zaini Md Zain, Iekhsan Othman, Sharifah Binti Syed Hassan

Abstract:

Meat quality is always subject to consumer scrutiny when purchasing from retail markets on mislabeling as fresh meat. Various physiological and biochemical changes influence the quality of meat. As a major component of muscle tissue, proteins play a major role in muscle foods. In meat industry, freezing is the most common form of storage of meat products. Repeated cycles of freezing and thawing are common in restaurants, kitchen, and retail outlets and can also occur during transportation or storage. Temperature fluctuation is responsible for physical, chemical, and biochemical changes. Repeated cycles of ‘freeze-thaw’ degrade the quality of meat by stimulating the lipid oxidation and surface discoloration. The shelf life of meat is usually determined by its appearance, texture, color, flavor, microbial activity, and nutritive value and is influenced by frozen storage and subsequent thawing. The main deterioration of frozen meat during storage is due to protein. Due to the large price differences between fresh and frozen–thawed meat, it is of great interest to consumer to know whether a meat product is truly fresh or not. Researchers have mainly focused on the reduction of moisture loss due to freezing and thawing cycles of meat. The water holding capacity (WHC) of muscle proteins and reduced water content are key quality parameters of meat that ultimately changes color and texture. However, there has been limited progress towards understanding the actual mechanisms behind the meat quality changes under the freeze–thaw cycles. Furthermore, effect of freeze-thaw process on integrity of proteins is ignored. In this paper, we have studied the effect of ‘freeze-thawing’ on physicochemical changes of chicken meat protein. We have assessed the quality of meat by pH, spectroscopic measurements, Western Blot. Our results showed that increase in freeze-thaw cycles causes changes in pH. Measurements of absorbance (UV-visible and IR) indicated the degradation of proteins. The expression of various proteins (CREB, AKT, MAPK, GAPDH, and phosphorylated forms) were performed using Western Blot. These results indicated the repeated cycles of freeze-thaw is responsible for deterioration of protein, thus causing decrease in nutritious value of meat. It damges the use of these products in Islamic Sharia.

Keywords: chicken meat, freeze-thaw, halal, protein, western blot

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Authors: Lucia Salvioni, Pietro Giorgio Lovaglio, Valerio Leoni, Miriam Colombo, Luisa Fiandra

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The involvement of plasmatic surfactant protein-D (SP-D) in pulmonary diseases has been long investigated, and over the last two years, more interest has been directed to determine its role as a marker of COVID-19. In this direction, several studies aimed to correlate pulmonary surfactant proteins with the clinical manifestations of the virus indicated SP-D as a prognostic biomarker of COVID-19 pneumonia severity. The present work has performed a retrospective study on a relatively large cohort of patients of Hospital Pio XI of Desio (Lombardia, Italy) with the aim to assess differences in the hematic SP-D concentrations among COVID-19 patients and healthy donors and the role of SP-D as a prognostic marker of severity and/or of mortality risk. The obtained results showed a significant difference in the mean of log SP-D levels between COVID-19 patients and healthy donors, so as between dead and survived patients. SP-D values were significantly higher for both hospitalized COVID-19 and dead patients, with threshold values of 150 and 250 ng/mL, respectively. SP-D levels at admission and increasing differences among follow-up and admission values resulted in the strongest significant risk factors of mortality. Therefore, this study demonstrated the role of SP-D as a predictive marker of SARS-CoV-2 infection and its outcome. A significant correlation of SP-D with patient mortality indicated that it is also a prognostic factor in terms of mortality, and its early detection should be considered to design adequate preventive treatments for COVID-19 patients.

Keywords: SARS-CoV-2 infection, COVID-19, surfactant protein-D (SP-D), mortality, biomarker

Procedia PDF Downloads 76

Authors: Aida Rafat, Ramazan Kahraman, Mert Atilhan

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The aggressive corrosion behavior of conventional amine solvents is one of main barriers against large scale commerizaliation of amine absorption process for carbon capture application. Novel CO2 absorbents that exhibit minimal corrosivity against operation conditions are essential to lower corrosion damage and control and ensure more robustness in the capture plant. This work investigated corrosion behavior of carbon steel CS1018 in various CO2 absrobent solvents. The tested solvents included the classical amines MEA, DEA and MDEA, piperazine activated solvents MEA/PZ, MDEA/PZ and MEA/MDEA/PZ as well as mixtures of MEA and Room Temperature Ionic Liquids RTIL, namely MEA/[C4MIM][BF4] and MEA/[C4MIM][Otf]. Electrochemical polarization technique was used to determine the system corrosiveness in terms of corrosion rate and polarization behavior. The process parameters of interest were CO2 loading and solution temperature. Electrochemical resulted showed corrosivity order of classical amines at 40°C is MDEA> MEA > DEA wherase at 80°C corrosivity ranking changes to MEA > DEA > MDEA. Corrosivity rankings were mainly governed by CO2 absorption capacity at the test temperature. Corrosivity ranking for activated amines at 80°C was MEA/PZ > MDEA/PZ > MEA/MDEA/PZ. Piperazine addition seemed to have a dual advanatge in terms of enhancing CO2 absorption capacity as well as nullifying corrosion. For MEA/RTIL mixtures, the preliminary results showed that the partial repalcement of aqueous phase in MEA solution by the more stable nonvolatile RTIL solvents reduced corrosion rates considerably.

Keywords: corrosion, amines, CO2 capture, piperazine, ionic liquids

Procedia PDF Downloads 460

Authors: Lucian Mocan, Flaviu Tabaran, Teodora Mocan, Cristian Matea, Cornel Iancu

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We present method of Gold Nanoparticle enhanced laser thermal ablation of HepG2 cells (Human hepatocellular liver carcinoma cell line), based on a simple gold nanoparticle carrier system, such as serum albumin (BSA), and demonstrate its selective therapeutic efficacy. Hyperspectral, contrast phase, and confocal microscopy combined immunochemical staining were used to demonstrate the selective internalization of HSA-GNPs via Gp60 receptors and the caveolin-mediated endocytosis inside HepG2 cells. We examined the ability of laser-activated carbon nanotubes to induce Hsp70 expression using confocal microscopy. Hep G2 cells heat-shocked (laser activated BSA-GNPs) to 42°C demonstrated an up-regulation of Hsp70 compared with control cells (BSA-GNPs treated cells without laser), which showed no detectable constitutive expression of Hsp70. We observed a time-dependent induction in Hsp70 expression in Hep G2 treated with BSA-GNPs and LASER irradiated. The post-irradiation apoptotic rate of HepG2 cells treated with HSA-GNPs ranged from 88.24% (for 50 mg/L) at 60 seconds, while at 30 minute the rate increased to 92.34% (50 mg/L). These unique results may represent a major step in liver cancer treatment using nanolocalized thermal ablation by laser heating.

Keywords: gold nanoparticles, liver cancer, albumin, laser irradiation

Procedia PDF Downloads 306