Search results for: immune modulation

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

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

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

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Authors: J. Adeppa, S. Samanta, O. K. Raina

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Fasciolosis is a widespread economically important parasitic infection throughout the world caused by Fasciola hepatica and F. gigantica. In order to identify novel immunogen conferring significant protection against fasciolosis, currently, research has been focused on the defined antigens viz. glutathione S-transferase, fatty acid binding protein, cathepsin-L, fluke hemoglobin, paramyosin, myosin and F. hepatica- Kunitz Type Molecule. Among various antigens, GST which plays a crucial role in detoxification processes, i.e. phase II defense mechanism of this parasite, has a unique position as a novel vaccine candidate and a drug target in the control of this disease. For producing the antigens in large quantities and their purification to complete homogeneity, the recombinant DNA technology has become an important tool to achieve this milestone. RT- PCR was carried out using F. gigantica total RNA as template, and an amplicon of 657 bp GST gene was obtained. TA cloning vector was used for cloning of this gene, and the presence of insert was confirmed by blue-white selection for recombinant colonies. Sequence analysis of the present isolate showed 99.1% sequence homology with the published sequence of the F. gigantica GST gene of cattle origin (accession no. AF112657), with six nucleotide changes at 72, 74, 423, 513, 549 and 627th bp found in the present isolate, causing an overall change of 4 amino acids. The 657 bp GST gene was cloned at BamH1 and HindIII restriction sites of the prokaryotic expression vector pPROEXHTb in frame with six histidine residues and expressed in E. coli DH5α. Recombinant protein was purified from the bacterial lysate under non-denaturing conditions by the process of sonication after lysozyme treatment and subjecting the soluble fraction of the bacterial lysate to Ni-NTA affinity chromatography. Western blotting with rabbit hyper-immune serum showed immuno-reactivity with 25 kDa recombinant GST. Recombinant protein detected F. gigantica experimental as well as field infection in buffaloes by dot-ELISA. However, cross-reactivity studies on Fasciola gigantica GST antigen are needed to evaluate the utility of this protein in the serodiagnosis of fasciolosis.

Keywords: fasciola gigantic, fasciola hepatica, GST, RT- PCR

Procedia PDF Downloads 187

Authors: Ecem Deniz Kırkpantur, Ozge Ecmel Onur, Tuba Cimilli Ozturk, Ebru Unal Akoglu

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Moyamoya disease is a unique chronic progressive cerebrovascular disease characterized by bilateral stenosis or occlusion of the arteries around the circle of Willis with prominent arterial collateral circulation. The occurrence of Moyamoya disease is related to immune, genetic and other factors. There is no curative treatment for Moyamoya disease. Secondary prevention for patients with symptomatic Moyamoya disease is largely centered on surgical revascularization techniques. We present here a 62-year old male presented with headache and vision loss for 2 days. He was previously diagnosed with hypertension and glaucoma. On physical examination, left eye movements were restricted medially, both eyes were hyperemic and their movements were painful. Other neurological and physical examination were normal. His vital signs and laboratory results were within normal limits. Computed tomography (CT) showed dilated vascular structures around both lateral ventricles and atherosclerotic changes inside the walls of internal carotid artery (ICA). Magnetic resonance imaging (MRI) and angiography (MRA) revealed dilated venous vascular structures around lateral ventricles and hyper-intense gliosis in periventricular white matter. Ischemic gliosis around the lateral ventricles were present in the Digital Subtracted Angiography (DSA). After the neurology, ophthalmology and neurosurgery consultation, the patient was diagnosed with Moyamoya disease, pulse steroid therapy was started for vision loss, and super-selective DSA was planned for further investigation. Moyamoya disease is a rare condition, but it can be an important cause of stroke in both children and adults. It generally affects anterior circulation, but posterior cerebral circulation may also be affected, as well. In the differential diagnosis of acute vision loss, occipital stroke related to Moyamoya disease should be considered. Direct and indirect surgical revascularization surgeries may be used to effectively revascularize affected brain areas, and have been shown to reduce risk of stroke.

Keywords: headache, Moyamoya disease, stroke, visual loss

Procedia PDF Downloads 267

Authors: Diya Kohli, Amalia Ardeljan, Lexi Frankel, Jose Garcia, Lokesh Manjani, Omar Rashid

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Gonorrhea is the second most common sexually transmitted disease (STDs) in the United States of America. Gonorrhea affects the urethra, rectum, or throat and the cervix in females. Lymphoma is a cancer of the immune network called the lymphatic system that includes the lymph nodes/glands, spleen, thymus gland, and bone marrow. Lymphoma can affect many organs in the body. When a lymphocyte develops a genetic mutation, it signals other cells into rapid proliferation that causes many mutated lymphocytes. Multiple studies have explored the incidence of cancer in people infected with STDs such as Gonorrhea. For instance, the studies conducted by Wang Y-C and Co., as well as Caini, S and Co. established a direct co-relationship between Gonorrhea infection and incidence of prostate cancer. We hypothesize that Gonorrhea infection also increases the incidence of Lymphoma in patients. This research study aimed to evaluate the correlation between Gonorrhea infection and the incidence of Lymphoma. The data for the research was provided by a Health Insurance Portability and Accountability Act (HIPAA) compliant national database. This database was utilized to evaluate patients infected with Gonorrhea versus the ones who were not infected to establish a correlation with the prevalence of Lymphoma using ICD-10 and ICD-9 codes. Access to the database was granted by the Holy Cross Health, Fort Lauderdale for academic research. Standard statistical methods were applied throughout. Between January 2010 and December 2019, the query was analyzed and resulted in 254 and 808 patients in both the infected and control group, respectively. The two groups were matched by Age Range and CCI score. The incidence of Lymphoma was 0.998% (254 patients out of 25455) in the Gonorrhea group (patients infected with Gonorrhea that was Lymphoma Positive) compared to 3.174% and 808 patients in the control group (Patients negative for Gonorrhea but with Lymphoma). This was statistically significant by a p-value < 2.210-16 with an OR= 0.431 (95% CI 0.381-0.487). The patients were then matched by antibiotic treatment to avoid treatment bias. The incidence of Lymphoma was 1.215% (82 patients out of 6,748) in the Gonorrhea group compared to 2.949% (199 patients out of 6748) in the control group. This was statistically significant by a p-value <5.410-10 with an OR= 0.468 (95% CI 0.367-0.596). The study shows a statistically significant correlation between Gonorrhea and a reduced incidence of Lymphoma. Further evaluation is recommended to assess the potential of Gonorrhea in reducing Lymphoma.

Keywords: gonorrhea, lymphoma, STDs, cancer, ICD

Procedia PDF Downloads 196

Authors: Osamede Asowata, Christo Pienaar, Johan Bekker

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Photovoltaic (PV) modules may become a trend for future PV systems because of their greater flexibility in distributed system expansion, easier installation due to their nature, and higher system-level energy harnessing capabilities under shaded or PV manufacturing mismatch conditions. This is as compared to the single or multi-string inverters. Novel residential scale PV arrays are commonly connected to the grid by a single DC–AC inverter connected to a series, parallel or series-parallel string of PV panels, or many small DC–AC inverters which connect one or two panels directly to the AC grid. With an increasing worldwide interest in sustainable energy production and use, there is renewed focus on the power electronic converter interface for DC energy sources. Three specific examples of such DC energy sources that will have a role in distributed generation and sustainable energy systems are the photovoltaic (PV) panel, the fuel cell stack, and batteries of various chemistries. A high-efficiency inverter using Metal Oxide Semiconductor Field-Effect Transistors (MOSFETs) for all active switches is presented for a non-isolated photovoltaic and AC-module applications. The proposed configuration features a high efficiency over a wide load range, low ground leakage current and low-output AC-current distortion with no need for split capacitors. The detailed power stage operating principles, pulse width modulation scheme, multilevel bootstrap power supply, and integrated gate drivers for the proposed inverter is described. Experimental results of a hardware prototype, show that not only are MOSFET efficient in the system, it also shows that the ground leakage current issues are alleviated in the proposed inverter and also a 98 % maximum associated driver circuit is achieved. This, in turn, provides the need for a possible photovoltaic panel switching technique. This will help to reduce the effect of cloud movements as well as improve the overall efficiency of the system.

Keywords: grid connected photovoltaic (PV), Matlab efficiency simulation, maximum power point tracking (MPPT), module integrated converters (MICs), multilevel converter, series connected converter

Procedia PDF Downloads 127

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

Procedia PDF Downloads 127

Authors: Maha Azzam, Mahmoud Gabr, Mahmoud Zakaria, Ayman Refaie, Amani Ismail, Sherry Khater, Sylvia Ashamallah, Mohamed Ghoniem

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Evidence was provided that human bone marrow-derived mesenhymal stem cells (HBM-MSCs) could be differentiated to form insulin-producing cells (IPCs). Transplantation of these cells was able to cure chemically-induced diabetes in nude mice. The efficacy of these cells to control diabetes in large animals was carried out to evaluate the sufficient number of cells needed/Kg body weight and to determine the functional longevity in vivo. Materials/Methods: Ten male mongrel dogs weighing 15-20 Kg were used in this study. Diabetes was chemically-induced in 7 dogs by a mixture of alloxan and streptozotocin. Three non-diabetic served as normal controls. Differentiated HBM-MSCs (5 million/Kg) were encapsulated in theracyte capsules and transplanted beneath the rectus sheath. Each dog received 2 capsules. One dog died 4 days postoperative from inhalation pneumonia. The remaining 6 dogs were followed up for 6-18 months. Results: Four dogs became normoglycemic within 6-8 weeks with normal glucose tolerance curves providing evidence that the transplanted cells were glucose-sensitive and insulin-responsive. In the remaining 2 dogs, fasting blood glucose was reduced but did not reach euglycemic levels. The sera of all transplanted dogs contained human insulin and c-peptide but negligible levels of canine insulin. When the HBM-MSCs loaded capsules were removed, rapid return of diabetic state was noted. The harvested capsules were examined by immunofluorescence. IPCs were seen and co-expression of with c-peptide was confirmed. Furthermore, all the pancreatic endocrine genes were expressed by the transplanted cells. Conclusions: This study provided evidence that theracyte capsules could protect the xenogenic HBM-MSCs from the host immune response. This is an important issue when clinical stem cell therapy is considered for definitive treatment for T1DM.

Keywords: diabetes, mesenchymal stem cells, dogs, Insulin-producing cells

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Authors: Rema Vazhappilly, Tapas Das

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Oleic acid is a cis unsaturated fatty acid and is known to be a partially essential fatty acid due to its limited endogenous synthesis during pregnancy and lactation. Previous studies have demonstrated the role of oleic acid in neuronal differentiation and brain phospholipid synthesis. These evidences indicate a major role for oleic acid in learning and memory. Interestingly, oleic acid has been shown to enhance hippocampal long term potentiation (LTP), the physiological correlate of long term synaptic plasticity. However the effect of oleic acid on short term synaptic plasticity has not been investigated. Short term potentiation (STP) is the physiological correlate of short term synaptic plasticity which is the key underlying molecular mechanism of short term memory and neuronal information processing. STP in the hippocampal CA1 region has been known to require the activation of N-methyl-D-aspartate receptors (NMDARs). The NMDAR dependent hippocampal STP as a potential mechanism for short term memory has been a subject of intense interest for the past few years. Therefore in the present study the effect of oleic acid on NMDAR dependent hippocampal STP was determined in mouse hippocampal slices (in vitro) using Multi-electrode array system. STP was induced by weak tetanic Stimulation (one train of 100 Hz stimulations for 0.1s) of the Schaffer collaterals of CA1 region of the hippocampus in slices treated with different concentrations of oleic acid in presence or absence of NMDAR antagonist D-AP5 (30 µM) . Oleic acid at 20 (mean increase in fEPSP amplitude = ~135 % Vs. Control = 100%; P<0.001) and 30 µM (mean increase in fEPSP amplitude = ~ 280% Vs. Control = 100%); P<0.001) significantly enhanced the STP following weak tetanic stimulation. Lower oleic acid concentrations at 10 µM did not modify the hippocampal STP induced by weak tetanic stimulation. The hippocampal STP induced by weak tetanic stimulation was completely blocked by the NMDA receptor antagonist D-AP5 (30µM) in both oleic acid and control treated hippocampal slices. This lead to the conclusion that the hippocampal STP elicited by weak tetanic stimulation and enhanced by oleic acid was NMDAR dependent. Together these findings suggest that oleic acid may enhance the short term memory and neuronal information processing through the modulation of NMDAR dependent hippocampal short-term synaptic plasticity. In conclusion this study suggests the possible role of oleic acid to prevent the short term memory loss and impaired neuronal function throughout development.

Keywords: oleic acid, short-term potentiation, memory, field excitatory post synaptic potentials, NMDA receptor

Procedia PDF Downloads 336

Authors: Liaqat Ali, Hubert E. Blum, Türkân Sakinc

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Enterococcus faecium is an emerging multidrug-resistant nosocomial pathogen increased dramatically worldwide and causing bacteremia, endocarditis, urinary tract and surgical site infections in immunocomprised patients. The capsular polysaccharides that contribute to pathogenesis through evasion of the host innate immune system are also involved in hindering leukocyte killing of enterococci. The gene cluster (enterococcal polysaccharide antigen) of E. faecalis encoding homologues of many genes involved in polysaccharide biosynthesis. We identified two putative loci with 22 kb and 19 kb which contained 11 genes encoding for glycosyltransferases (GTFs); this was confirmed by using genome comparison of already sequenced strains that has no homology to known capsule genes and the epa-locus. The polysaccharide-conjugate vaccines have rapidly emerged as a suitable strategy to combat different pathogenic bacteria, therefore, we investigated a polysaccharide biosynthesis CapD protein in E. faecium contains 336 amino acids and had putative function for N-linked glycosylation. The deletion/knock-out capD mutant was constructed and complemented by homologues recombination method and confirmed by using PCR and sequencing. For further characterization and functional analysis, in-vitro cell culture and in-vivo a mouse infection models were used. Our ΔcapD mutant shows a strong hydrophobicity and all strains exhibited biofilm production. Subsequently, the opsonic activity was tested in an opsonophagocytic assay which shows increased in mutant compared complemented and wild type strains but more than two fold decreased in colonization and adherence was seen on surface of uroepithelial cells. However, a significant higher bacterial colonialization was observed in capD mutant during animal bacteremia infection. Unlike other polysaccharides biosynthesis proteins, CapD does not seems to be a major virulence factor in enterococci but further experiments and attention is needed to clarify its function, exact mechanism and involvement in pathogenesis of enteroccocal nosocomial infections eventually to develop a vaccine/ or targeted therapy.

Keywords: E. faecium, pathogenesis, polysaccharides, biofilm formation

Procedia PDF Downloads 334

Authors: Chyuan Haur Kao, Hsiang Chen, Yu Sheng Tsai, Chen Hao Hung, Yu Shan Lee

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Dielectric electrolyte-insulator-semiconductor sensing membranes-based biosensors have been intensively investigated because of their simple fabrication, low cost, and fast response. However, to enhance their sensing performance, it is worthwhile to explore alternative materials, distinct processes, and novel treatments. An ISFET can be viewed as a variation of MOSFET with the dielectric oxide layer as the sensing membrane. Then, modulation on the work function of the gate caused by electrolytes in various ion concentrations could be used to calculate the ion concentrations. Recently, owing to the advancement of CMOS technology, some high dielectric materials substrates as the sensing membranes of electrolyte-insulator-semiconductor (EIS) structures. The EIS with a stacked-layer of SiO₂ layer between the sensing membrane and the silicon substrate exhibited a high pH sensitivity and good long-term stability. IGZO is a wide-bandgap (~3.15eV) semiconductor of the III-VI semiconductor group with several preferable properties, including good transparency, high electron mobility, wide band gap, and comparable with CMOS technology. IGZO was sputtered by reactive radio frequency (RF) on a p-type silicon wafer with various gas ratios of Ar:O₂ and was treated with rapid thermal annealing in O₂ ambient. The sensing performance, including sensitivity, hysteresis, and drift rate was measured and XRD, XPS, and AFM analyses were also used to study the material properties of the IGZO membrane. Moreover, IGZO was used as a sensing membrane in dielectric EIS bio-sensor structures. In addition to traditional pH sensing capability, detection for concentrations of Na+, K+, urea, glucose, and creatinine was performed. Moreover, post rapid thermal annealing (RTA) treatment was confirmed to improve the material properties and enhance the multi-analyte sensing capability for various ions or chemicals in solutions. In this study, the IGZO sensing membrane with annealing in O₂ ambient exhibited a higher sensitivity, higher linearity, higher H+ selectivity, lower hysteresis voltage and lower drift rate. Results indicate that the IGZO dielectric sensing membrane on the EIS structure is promising for future bio-medical device applications.

Keywords: dielectric sensing membrane, IGZO, hydrogen ion, plasma, rapid thermal annealing

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Authors: Dinka Fikadu, Mulugeta Shegaze

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Background: Currently in health facility, provider-initiated human immunodeficiency virus testing is the key entry point to prevention, care, treatment and support services, but most people remains unaware of their HIV status due to various reasons. In many high-prevalence countries, fewer than one in ten people with HIV are aware of their HIV status. HIV, the virus that causes AIDS, “acquired immunodeficiency syndrome, "has become one of the world’s most serious health and development challenges. Reaching individuals with HIV who do not know their serostatus is a global public health priority. Objective: To assess utilization of provider initiated HIV testing and counseling and associated factors among adult outpatient department patients. Methods: Health facility based cross sectional study was conducted among 392 adult outpatient department patients in Wonchi woreda from February 24 to March 24 /2013. The study participant was recruited patients from all adult outpatient department patients of all four public health facilities of wonchi woreda using systematic sampling. A structured interviewer administered questionnaire was used to elicit all important variables from the study participants and multiple logistic regression analysis was used. Result: A total of 371 adult outpatient department patients aged between 15 to 64 years were actively participated in the study and 291(78.4%) of them utilized provider initiated HIV testing and counseling and 80(21.6%) of them refused. Knowledge on HIV is low in the study population; majority of the participants didn’t have comprehensive knowledge (64.7%) and (35.3%) fail to reject misconception about means of HIV transmission and prevention. Utilization of provider-initiated HIV testing and counseling were associated with divorced/widowed marital status[AOR (95%CI) = 0.32(0.15, 0.69)], being male sex [AOR (95%CI) =1.81(1.01, 3.24)], having comprehensive knowledge on HIV [AOR (95%CI) =0.408(0.220,0.759)],having awareness about provider initiated HIV testing and counseling [AOR(95%CI) =2.89(1.48,5.66)] and receiving test on HIV before[AOR (95%CI)=4.15(2.30, 7.47)]. Conclusion: Utilization of provider initiated HIV testing and counseling among adult outpatient departments in wonchi woreda public health facility was [(78.4%)].Strengthening health information through mass media and peer education on HIV to address barrier to testing in the community such as low awareness on PITC, to increase up take of PITC among adult OPD patients.

Keywords: utilization, human immune deficiency, testing, provider, initiate

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Authors: L. L. Tundisi, A. Pessoa Jr., E. B. Tambourgi, E. Silveira, P. G. Mazzola

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L-asparaginase (L-ASNase) is the gold standard treatment for acute lymphoblastic leukemia that mainly affects pediatric patients; treatment increases survival from 20% to 90%. The characterization of other L-Asparaginases, apart from the most used from Escherichia coli and Erwinia chrysanthemi, has been reported, but the choice of the most appropriate is still under debate. This choice should be based on its pharmacokinetics, immune hypersensitivity, doses, prices, pharmacodynamics. The main factors influencing the antileukemic activity of ASNase are enzymatic activity, Km, glutaminase activity, clearance of the enzyme and development of resistance. However, most of the commercialized enzyme present an intrinsic glutaminase activity, which is responsible for some side effects. In this study, glutaminase free asparaginase produced from Aspergillus oryzae was precipitated in different percentages of ethanol (0–80%), until optimum ethanol concentration of 60% (w/w) was found. Following, precipitation of crude L-ASNase was performed in a single step, using 60% (w/w) ethanol, under constant agitation and temperature. It presented activity of 135.45 U/mg and after gel filtration chromatography with Sephadex G-the enzymatic activity was 322.02 U/mg. The apparent molecular mass of the purified L-ASNase fraction was estimated by 10% SDS-PAGE. Proteins were stained with Coomassie Brilliant Blue R-250. The molar mass range was from 10 kDa to 250 kDa. L-ASNase from Aspergillus oryzae was characterized aiming possible therapeutic use. Four different buffers (phosphate-citrate buffer pH 2.6 to 5.8; phosphate buffer pH 5.8 to 7.4; Tris - HCl pH 7.4 to 9.0; and carbonate buffer pH 9.8 to 10.6) were used to measure the optimum pH for L-ASNase activity. The optimum temperature for enzyme activity was measured at optimal pH conditions (Tris-HCl and phosphate buffer, pH 7.4) at different temperatures ranging from 5 to 55°C. All activities were calculated by quantifying the free ammonia, using the Nessler reagent. The kinetic parameters calculation, e.g. Michaelis-Menten constant (Km), maximum velocity (Vmax) and Hills coefficient (n), were performed by incubating the enzyme in different concentrations of the substrate at optimum conditions of pH and fitted on Hill’s equation. This glutaminase free asparaginase showed a low Km (3.39 mM and 3.81 mM) and enzymatic activity of 135.45 U/mg after precipitation with ethanol. After gel filtration chromatography it rose to 322.02 U/mg. Optimum activity was found between pH 5.8 - 9.0, best activity results with phosphate buffer pH 7.4 and Tris-HCl pH 7.4 and showed activity from 5°C to 55°C. These results indicate that L-ASNase from A. oryzae has the potential for human use.

Keywords: biopharmaceuticals, bioprocessing, bioproducts, biotechnology, enzyme activity, ethanol precipitation

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Authors: Akbar Haghzadeh Saraskanroud, Amir Hossein Yahyavi Zanjani, Niloofar Kargar, Hanieh Ahrabi

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Cervical cancer refers to an unusual growth of cells in the cervix. The cervix is the lower part of the uterus, which connects to the vagina. Various risk factors such as human papillomavirus (HPV), having a weakened immune system, smoking or breathing in secondhand smoke, reproductive factors, and obesity play important roles in causing most cervical cancers. When cervical cancer happens, surgery is often the first treatment option to remove it. Other treatments might include chemotherapy and targeted therapy medicines. Radiation therapy with high-energy photon beams also may be used. Sometimes combined treatment, including radiation with low-dose chemotherapy, was applied. Intracavitary brachytherapy is an integral part of radiotherapy for locally advanced gynecologic malignancies such as cervical cancer. In the treatment of cervical cancer, there are different tools for doing brachytherapy. Two combinations of different applicators for this purpose are Tandem and Ovoid and Tandem and Ring. This study evaluated the dose differences between these two methods in the organs at risk of the rectum, sigmoid, and bladder. In this study, the treatment planswere simulated by the Oncentra treatment planning system and Tandem, Ovid, and Rings of different sizes. CT scan images of 23 patients were treated with HDR_BT Elekta Flexitron system were used for this study. Contouring of HR-CTV, rectum and bladder was performed for all patients. Then, the received dose of 0.1 and 0.2cc volumes of organs at risk were obtained and compared for these two methods: T-Ovoid and T-Ring. By doing investigations and dose measurements of points A and B and the volumes specified by ICRU, it seems that when comparing ring and ovoid to tandem and ovoid, the total dose to the rectum was lower by about 11%, and the bladder was 7%. In the case of HR CTV, this comparison showed that this ratio is about 7% better. Figure 1 shows the amount of decrease in rectum dose in the T-Ring method compared to T-Ovoid. Figure 2 indicates the amount of decrease in bladder dose in the T-Ring method compared to T-Ovoid. Finally, figure 3 illustrates the amount of HR-CTV coverage in the T-Ring method compared to the T-Ovoid.

Keywords: cervical cancer, brachytherapy, rectum, tandem and ovoid, tandem and ring.

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Authors: Junaid Mahmood Alam, Sana Anwar, Sarah Sughra Asghar

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Chronic hepatitis or Hepatitis C viral (HCV) infection has been identified as one of the factors that could elicit autoimmune disease resulting in the development of auto-antibodies. Furthermore, HCV is implicated in contravening of forbearance to antigens, therefore, inciting auto-reactivity. In this regard, several near and past studies noted the prevalence of thyroid dysfunction and production of anti-thyroid antibodies (ATAb) such as anti-thyroid peroxidase (AntiTPO) and anti-thyroglobulin (AntiTG) in patients with HCV. Likewise, one of the etiologies of augmentation of thyroid disease is basically interferon therapy for HCV infections, for which a number of autoimmune diseases have been noted including Grave’s disease, Hishimoto thyroiditis. A prospectively case-control study was therefore carried out at department of clinical biochemistry lab services and chemical pathology in collaboration with department of clinical microbiology, at Liaquat National Hospital and Medical College, Karachi Pakistan for the period January 2015 to December 2017. Two control groups were inducted for comparison purpose, control group 1 = without HCV infection and with thyroid disorders (n = 20), control group 2 = with HCV infection and without thyroid disorders (n = 20), whereas HCV infected were n = 40 where more than half were noted to be positive for either of HCV IgG and Ag. In HCV group, patients with existing sub-clinical hypothyroidism and clinical hyperthyroidism were less than 5%. Analysis showed the presence of AntiTG in 12 HCV patients (30%), AntiTPO in 15 (37.5%) and both AntiTG and antiTPO in 10 patients (25%). Only 3 patients were found with the history of anti-thyroid auto-antibodies (7.5%) and one with parents and relatives with auto-immune disorders (2.5%). Patients that remained untreated were 12 (30%), under treatment 18 (45%) and with complete-course of treatment 10 (25%). As per review of the literature, meta-analysis of evident data and cross-sectional studies of selective cohorts (as studied in presented research), thyroid connection is designated as one of the most recurrent endocrine ailment associated with chronic HCV infection. Moreover, it also represents an extrahepatic disease in the continuum of HCV syndrome. In conclusion, HCV patients were more likely to encompass thyroid disorders especially related to development of either of ATAb or both antiTG and AntiTPO.

Keywords: Hepatitis C viral (HCV) infection, anti-thyroid antibodies, anti-thyroid peroxidase antibodies, anti-thyroglobulin antibodies

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Authors: Seunghwan Seo, Woo-Seok Lee, Ji-Sun Shin, Young Kyoung Rhee, Chang-Won Cho, Hee-Do Hong, Kyung-Tae Lee

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Doenjang, known as traditional Korean food, is product of a natural mixed fermentation process carried out by lactic acid bacteria (LAB). Lactobacillus sakei K040706 (K040706) has been accepted as the most populous LAB in over ripened doenjang. Recently, we reported the immunostimulatory effects of K040706 in RAW 264.7 macrophages and in a cyclophosphamide-induced mouse model. In this study, we investigated the ameliorative effects of K040706 in a dextran sulfate sodium (DSS)-induced colitis mouse model. We induced colitis using DSS in 5-week-ICR mice over 14 days with or without 0.1, 1 g/kg/day K040706 orally. The body weight, stool consistency, and gross bleeding were recorded for determination of the disease activity index (DAI). At the end of treatment, animals were sacrificed and colonic tissues were collected and subjected to histological experiments and myeloperoxidase (MPO) accumulation, cytokine determination, qRT-PCR and Western blot analysis. Results showed that K040706 significantly attenuated DSS-induced DAI score, shortening of colon length, enlargement of spleen and immune cell infiltrations into colonic tissues. Histological examinations indicated that K040706 suppressed edema, mucosal damage, and the loss of crypts induced by DSS. These results were correlated with the restoration of tight junction protein expression, such as, ZO-1 and occludin in K040706-treated mice. Moreover, K040706 reduced the abnormal secretions and mRNA expressions of pro-inflammatory mediators, such as nitric oxide (NO), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and interleukin-6 (IL-6). DSS-induced mRNA expression of intercellular adhesion molecule (ICAM) and vascular cell adhesion molecule (VCAM) in colonic tissues was also downregulated by K040706 treatment. Furthermore, K040706 suppressed the protein and mRNA expression of toll-like receptor 4 (TLR4) and phosphorylation of NF-κB and signal transducer and activator of transcription 3 (STAT3). These results suggest that K040706 has an anti-colitic effect by inhibition of intestinal inflammatory responses in DSS-induced colitic mice.

Keywords: Lactobacillus sakei, NF-κB, STAT3, ulcerative colitis

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Authors: Vijaya Madhuri Devraj, Swarnalatha Guditi, Kiran Kumar Bokara, Gangadhar Taduri

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Cell-based strategies may open therapeutic approaches that promote tolerance through manipulation of macrophages to increase long-term transplant survival rates and minimize side effects of the current immune suppressive regimens. The aim of the present study was, therefore, to test and compare the therapeutic potential of MSC and Tregs on macrophage polarization to develop an alternate cell-based treatment option in kidney transplantation. In the current protocol, macrophages from kidney transplant recipients with graft dysfunction were co-cultured with MSCs and Treg cells with and without cell-cell contact on transwell plates, further to quantitatively assess macrophage polarization in response to MSC and Treg treatment over time, M1 and M2 cell surface markers were used. Additionally, multiple soluble analytes were analyzed in cell supernatant by using bead-based immunoassays. Furthermore, to confirm our findings, gene expression analysis was done. MSCs induced the formation of M2 macrophages more than Tregs when macrophages M0 were cultured in transwell without cell contact. From this, we deduced the mechanism that soluble factors present in the MSCs condition media are involved in skewing of macrophages towards type 2 macrophages; similarly, in co-culture with cell-cell contact, MSCs resulted in more M2 type macrophages than Tregs. And an important finding of this study is the combination of both MSC-Treg showed significantly effective and consistent results in both with and without cell contact setups. Hence, it is suggestive to prefer MSCs over Tregs for secretome-based therapy and a combination of both for either therapy for effective transplantation outcomes. Our findings underline a key role of Tregs and MSCs in promoting macrophage polarization towards anti-inflammatory type. The study has great importance in prolongation of allograft and patient survival without any rejection by cell-based therapy, which induce self-tolerance and controlling infection.

Keywords: graft rejection, graft tolerance, macrophage polarization, mesenchymal stem cells, regulatory T cells, transplant immunology

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Authors: Ieva Bruzauskaite, Jovile Raudoniute, Karina Poliakovaite, Danguole Zabulyte, Daiva Bironaite, Ruta Aldonyte

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Introduction: Air pollution is a growing global problem which has been shown to be responsible for various adverse health outcomes. Immunotoxicity, such as dysregulated inflammation, has been proposed as one of the main mechanisms in air pollution-associated diseases. Chronic obstructive pulmonary disease (COPD) is among major morbidity and mortality causes worldwide and is characterized by persistent airflow limitation caused by the small airways disease (obstructive bronchiolitis) and irreversible parenchymal destruction (emphysema). Exact pathways explaining the air pollution induced and mediated disease states are still not clear. However, modern societies understand dangers of polluted air, seek to mitigate such effects and are in need for reliable biomarkers of air pollution. We hypothesise that post-translational modifications of structural proteins, e.g. citrullination, might be a good candidate biomarker. Thus, we have designed this study, where mice were exposed to diesel exhaust and the ongoing protein modifications and inflammation in lungs and other tissues were assessed. Materials And Methods: To assess the effects of diesel exhaust a in vivo study was designed. Mice (n=10) were subjected to everyday 2-hour exposure to diesel exhaust for 14 days. Control mice were treated the same way without diesel exhaust. The effects within lung and other tissues were assessed by immunohistochemistry of formalin-fixed and paraffin-embedded tissues. Levels of inflammation and citrullination related markers were investigated. Levels of parenchymal damage were also measured. Results: In vivo study corroborates our own data from in vitro and reveals diesel exhaust initiated inflammatory shift and modulation of lung peptidyl arginine deiminase 4 (PAD4), citrullination associated enzyme, levels. In addition, high levels of citrulline were observed in exposed lung tissue sections co-localising with increased parenchymal destruction. Conclusions: Subacute exposure to diesel exhaust renders mice lungs inflammatory and modifies certain structural proteins. Such structural changes of proteins may pave a pathways to lost/gain function of affected molecules and also propagate autoimmune processes within the lung and systemically.

Keywords: air pollution, citrullination, in vivo, lungs

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Authors: Victor Lopez-Richard, Rafael Schio Wengenroth Silva, Fabian Hartmann

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Memcomputing is a computational paradigm that combines information processing and storage on the same physical platform. Key elements for this topic are devices with an inherent memory, such as memristors, memcapacitors, and meminductors. Despite the widespread emergence of memory effects in various solid systems, a clear understanding of the basic microscopic mechanisms that trigger them is still a puzzling task. We report basic ingredients of the theory of solid-state transport, intrinsic to a wide range of mechanisms, as sufficient conditions for a memristive response that points to the natural emergence of memory. This emergence should be discernible under an adequate set of driving inputs, as highlighted by our theoretical prediction and general common trends can be thus listed that become a rule and not the exception, with contrasting signatures according to symmetry constraints, either built-in or induced by external factors at the microscopic level. Explicit analytical figures of merit for the memory modulation of the conductance are presented, unveiling very concise and accessible correlations between general intrinsic microscopic parameters such as relaxation times, activation energies, and efficiencies (encountered throughout various fields in Physics) with external drives: voltage pulses, temperature, illumination, etc. These building blocks of memory can be extended to a vast universe of materials and devices, with combinations of parallel and independent transport channels, providing an efficient and unified physical explanation for a wide class of resistive memory devices that have emerged in recent years. Its simplicity and practicality have also allowed a direct correlation with reported experimental observations with the potential of pointing out the optimal driving configurations. The main methodological tools used to combine three quantum transport approaches, Drude-like model, Landauer-Buttiker formalism, and field-effect transistor emulators, with the microscopic characterization of nonequilibrium dynamics. Both qualitative and quantitative agreements with available experimental responses are provided for validating the main hypothesis. This analysis also shades light on the basic universality of complex natural impedances of systems out of equilibrium and might help pave the way for new trends in the area of memory formation as well as in its technological applications.

Keywords: memories, memdevices, memristors, nonequilibrium states

Procedia PDF Downloads 99

Authors: Benjamin Bressolette, S´ebastien Denjean, Vincent Roussarie, Mitsuko Aramaki, Sølvi Ystad, Richard Kronland-Martinet

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New sensors and technologies – such as microphones, touchscreens or infrared sensors – are currently making their appearance in the automotive sector, introducing new kinds of Human-Machine Interfaces (HMIs). The interactions with such tools might be cognitively expensive, thus unsuitable for driving tasks. It could for instance be dangerous to use touchscreens with a visual feedback while driving, as it distracts the driver’s visual attention away from the road. Furthermore, new technologies in car cockpits modify the interactions of the users with the central system. In particular, touchscreens are preferred to arrays of buttons for space improvement and design purposes. However, the buttons’ tactile feedback is no more available to the driver, which makes such interfaces more difficult to manipulate while driving. Gestures combined with an auditory feedback might therefore constitute an interesting alternative to interact with the HMI. Indeed, gestures can be performed without vision, which means that the driver’s visual attention can be totally dedicated to the driving task. In fact, the auditory feedback can both inform the driver with respect to the task performed on the interface and on the performed gesture, which might constitute a possible solution to the lack of tactile information. As audition is a relatively unused sense in automotive contexts, gesture sonification can contribute to reducing the cognitive load thanks to the proposed multisensory exploitation. Our approach consists in using a virtual object (VO) to sonify the consequences of the gesture rather than the gesture itself. This approach is motivated by an ecological point of view: Gestures do not make sound, but their consequences do. In this experiment, the aim was to identify efficient sound strategies, to transmit dynamic information of VOs to users through sound. The swipe gesture was chosen for this purpose, as it is commonly used in current and new interfaces. We chose two VO parameters to sonify, the hand-VO distance and the VO velocity. Two kinds of sound parameters can be chosen to sonify the VO behavior: Spectral or temporal parameters. Pitch and brightness were tested as spectral parameters, and amplitude modulation as a temporal parameter. Performances showed a positive effect of sound compared to a no-sound situation, revealing the usefulness of sounds to accomplish the task.

Keywords: auditory feedback, gesture sonification, sound perception, virtual object

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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: 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 363

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 186

Authors: Daniel Bülz, Franziska Lüttich, Sreetama Banerjee, Georgeta Salvan, Dietrich R. T. Zahn

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For the development of electronics, organic semiconductors are of great interest due to their adjustable optical and electrical properties. Especially for spintronic applications they are interesting because of their weak spin scattering, which leads to longer spin life times compared to inorganic semiconductors. It was shown that some organic materials change their resistance if an external magnetic field is applied. Pentacene is one of the materials which exhibit the so called photoinduced magnetoresistance which results in a modulation of photocurrent when varying the external magnetic field. Also the soluble derivate of pentacene, the 6,13-bis (triisopropylsilylethynyl)-pentacene (TIPS-pentacene) exhibits the same negative magnetoresistance. Aiming for simpler fabrication processes, in this work, we compare TIPS-pentacene organic field effect transistors (OFETs) made from solution with those fabricated by thermal evaporation. Because of the different processing, the TIPS-pentacene thin films exhibit different morphologies in terms of crystal size and homogeneity of the substrate coverage. On the other hand, the interface treatment is known to have a high influence on the threshold voltage, eliminating trap states of silicon oxide at the gate electrode and thereby changing the electrical switching response of the transistors. Therefore, we investigate the influence of interface treatment using octadecyltrichlorosilane (OTS) or using a simple cleaning procedure with acetone, ethanol, and deionized water. The transistors consist of a prestructured OFET substrates including gate, source, and drain electrodes, on top of which TIPS-pentacene dissolved in a mixture of tetralin and toluene is deposited by drop-, spray-, and spin-coating. Thereafter we keep the sample for one hour at a temperature of 60 °C. For the transistor fabrication by thermal evaporation the prestructured OFET substrates are also kept at a temperature of 60 °C during deposition with a rate of 0.3 nm/min and at a pressure below 10-6 mbar. The OFETs are characterized by means of optical microscopy in order to determine the overall quality of the sample, i.e. crystal size and coverage of the channel region. The output and transfer characteristics are measured in the dark and under illumination provided by a white light LED in the spectral range from 450 nm to 650 nm with a power density of (8±2) mW/cm2.

Keywords: organic field effect transistors, solution processed, surface treatment, TIPS-pentacene

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Authors: Govinda Pathak

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In modern agriculture, the sustainable enhancement of crop productivity while minimizing environmental impacts remains a paramount challenge. Plant Growth Promoting Rhizobacteria (PGPR) have emerged as a promising solution to address this challenge. The rhizosphere, the dynamic interface between plant roots and soil, hosts intricate microbial interactions crucial for plant health and nutrient acquisition. PGPR, a subset of rhizospheric microorganisms, exhibit multifaceted beneficial effects on plants. Their abilities to stimulate growth, confer stress tolerance, enhance nutrient availability, and suppress pathogens make them invaluable contributors to sustainable agriculture. This work examines the pivotal role of free living nitrogen fixer in optimizing agricultural practices. We delve into the intricate mechanisms underlying PGPR-mediated plant-microbe interactions, encompassing quorum sensing, root exudate modulation, and signaling molecule exchange. Furthermore, we explore the diverse strategies employed by PGPR to enhance plant resilience against abiotic stresses such as drought, salinity, and metal toxicity. Additionally, we highlight the role of PGPR in augmenting nutrient acquisition and soil fertility through mechanisms such as nitrogen fixation, phosphorus solubilization, and mineral mobilization. Furthermore, we discuss the potential of PGPR in minimizing the reliance on chemical fertilizers and pesticides, thereby contributing to environmentally friendly agriculture. However, harnessing the full potential of PGPR requires a comprehensive understanding of their interactions with host plants and the surrounding microbial community. We also address challenges associated with PGPR application, including formulation, compatibility, and field efficacy. As the quest for sustainable agriculture intensifies, harnessing the remarkable attributes of PGPR offers a holistic approach to propel agricultural productivity while maintaining ecological balance. This work underscores the promising prospect of free living nitrogen fixer as a panacea for addressing critical agricultural challenges regarding chemical urea in an era of sustainable and resilient food production.

Keywords: PGPR, nitrogen fixer, quorum sensing, Rhizobacteria, pesticides

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Authors: Seyedeh Banafsheh Bagheri Marzouni, Sona Rostampour Yasouri

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Salmonella is one of the most important common pathogens between humans and animals worldwide. Globally, the prevalence of the disease in humans is due to the consumption of food contaminated with animal-derived Salmonella. These foods include eggs, red meat, chicken, and milk. Contamination of chicken and its products with Salmonella may occur at any stage of the chicken processing chain. Salmonella infection is usually not fatal. However, its occurrence is considered dangerous in some individuals, such as infants, children, the elderly, pregnant women, or individuals with weakened immune systems. If Salmonella infection enters the bloodstream, the possibility of contamination of tissues throughout the body will arise. Therefore, determining the potential risk of Salmonella at various stages is essential from the perspective of consumers and public health. The aim of this study is to isolate and identify Salmonella from chicken samples distributed in the Tehran market using the Gold standard culture method and PCR techniques based on specific genes, invA and ent. During the years 2022-2023, sampling was performed using swabs from the liver and intestinal contents of distributed chickens in the Tehran province, with a total of 120 samples taken under aseptic conditions. The samples were initially enriched in buffered peptone water (BPW) for pre-enrichment overnight. Then, the samples were incubated in selective enrichment media, including TT broth and RVS medium, at temperatures of 37°C and 42°C, respectively, for 18 to 24 hours. Organisms that grew in the liquid medium and produced turbidity were transferred to selective media (XLD and BGA) and incubated overnight at 37°C for isolation. Suspicious Salmonella colonies were selected for DNA extraction, and PCR technique was performed using specific primers that targeted the invA and ent genes in Salmonella. The results indicated that 94 samples were Salmonella using the PCR technique. Of these, 71 samples were positive based on the invA gene, and 23 samples were positive based on the ent gene. Although the culture technique is the Gold standard, PCR is a faster and more accurate method. Rapid detection through PCR can enable the identification of Salmonella contamination in food items and the implementation of necessary measures for disease control and prevention.

Keywords: culture, PCR, salmonella spp, salmonella enteritidis

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Authors: Walid Wahballah, Taher Bazan, Fawzy Eltohamy

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One of the important functions of the satellite attitude control system is to provide the required pointing accuracy and attitude stability for optical remote sensing satellites to achieve good image quality. Although offering noise reduction and increased sensitivity, time delay and integration (TDI) charge coupled devices (CCDs) utilized in high-resolution satellites (HRS) are prone to introduce large amounts of pixel smear due to the instability of the line of sight. During on-orbit imaging, as a result of the Earth’s rotation and the satellite platform instability, the moving direction of the TDI-CCD linear array and the imaging direction of the camera become different. The speed of the image moving on the image plane (focal plane) represents the image motion velocity whereas the angle between the two directions is known as the drift angle (β). The drift angle occurs due to the rotation of the earth around its axis during satellite imaging; affecting the geometric accuracy and, consequently, causing image quality degradation. Therefore, the image motion velocity vector and the drift angle are two important factors used in the assessment of the image quality of TDI-CCD based optical remote sensing satellites. A model for estimating the image motion velocity and the drift angle in HRS is derived. The six satellite attitude control parameters represented in the derived model are the (roll angle φ, pitch angle θ, yaw angle ψ, roll angular velocity φ֗, pitch angular velocity θ֗ and yaw angular velocity ψ֗ ). The influence of these attitude parameters on the image quality is analyzed by establishing a relationship between the image motion velocity vector, drift angle and the six satellite attitude parameters. The influence of the satellite attitude parameters on the image quality is assessed by the presented model in terms of modulation transfer function (MTF) in both cross- and along-track directions. Three different cases representing the effect of pointing accuracy (φ, θ, ψ) bias are considered using four different sets of pointing accuracy typical values, while the satellite attitude stability parameters are ideal. In the same manner, the influence of satellite attitude stability (φ֗, θ֗, ψ֗) on image quality is also analysed for ideal pointing accuracy parameters. The results reveal that cross-track image quality is influenced seriously by the yaw angle bias and the roll angular velocity bias, while along-track image quality is influenced only by the pitch angular velocity bias.

Keywords: high-resolution satellites, pointing accuracy, attitude stability, TDI-CCD, smear, MTF

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Authors: Yemane Tadesse Gebreslassie, Fisseha Guesh Gebremeskel

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

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

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Authors: M. Bolacali

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The aim of the study is to determine the effect of immunocastration vaccine administration at different doses on fattening performance of feedlot Holstein bulls. Bopriva® is a vaccine that stimulates the animals' own immune system to produce specific antibodies against gonadotropin releasing factor (GnRF). Ninety four Holstein male calves (309.5 ± 2.58 kg body live weight and 267 d-old) assigned to the 4 treatments. Control group; 1 mL of 0.9% saline solution was subcutaneously injected to intact bulls on 1st and 60th days of the feedlot as placebo. On the same days of the feedlot, Bopriva® at two doses of 1 mL and 1 mL for Trial-1 group, 1.5 mL, and 1.5 mL for Trial-2 group, 1.5 mL, and 1 mL for Trial-3 group were subcutaneously injected to bulls. The study was conducted in a private establishment in the Sirvan district of Siirt province and lasted 180 days. The animals were weighed at the beginning of fattening and at 30-day intervals to determine their live weights at various periods. The statistical analysis for normal distribution data of the treatment groups was carried out with the general linear model procedure of SPSS software. The fattening initial live weight in Control, Trial-1, Trial-2 and Trial-3 groups was respectively 309.21, 306.62, 312.11, and 315.39 kg. The fattening final live weight was respectively 560.88, 536.67, 548.56, and 548.25 kg. The daily live weight gain during the trial was respectively 1.40, 1.28, 1.31, and 1.29 kg/day. The cold carcass yield was respectively 51.59%, 50.32%, 50.85%, and 50.77%. Immunocastration vaccine administration at different doses did not affect the live weights and cold carcass yields of Holstein male calves reared under intensive conditions (P > 0.05). However, it was determined to reduce fattening performance between 61-120 days (P < 0.05) and 1-180 days (P < 0.01). In addition, it was determined that the best performance among the vaccine-treated groups occurred in the group administered a 1.5 mL of vaccine on the 1st and 60th study days. In animals, castration is used to control fertility, aggressive and sexual behaviors. As a result, the fact that stress is induced by physical castration in animals and active immunization against GnRF maintains performance by maximizing welfare in bulls improves carcass and meat quality and controls unwanted sexual and aggressive behavior. Considering such features, it may be suggested that immunocastration vaccine with Bopriva® can be administered as a 1.5 mL dose on the 1st and 60th days of the fattening period in Holstein bulls.

Keywords: anti-GnRF, fattening, growth, immunocastration

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Authors: Sunghee Nam

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

Keywords: optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi optimization of culture conditions of paecilomyces tenuipes, entomopathogenic fungi silkworm larva, bombyx mori

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Authors: Omar Nouri, Wafa Mnejja, Nejla Fourati, Fatma Dhouib, Wicem Siala, Ilhem Charfeddine, Afef Khanfir, Jamel Daoud

Abstract:

Background and objective: Induction chemotherapy (IC) followed by concomitant chemo radiotherapy with intensity modulated radiation (IMRT) technique is actually the recommended treatment modality for locally advanced nasopharyngeal carcinomas (NPC). The aim of this study was to evaluate the prognostic factors predicting loco regional relapse with this new treatment protocol. Patients and methods: A retrospective study of 52 patients with NPC treated between June 2016 and July 2019. All patients received IC according to the protocol of the Head and Neck Radiotherapy Oncology Group (Gortec) NPC 2006 (3 TPF courses) followed by concomitant chemo radiotherapy with weekly cisplatin (40 mg / m2). Patients received IMRT with integrated simultaneous boost (SIB) of 33 daily fractions at a dose of 69.96 Gy for high-risk volume, 60 Gy for intermediate risk volume and 54 Gy for low-risk volume. Median age was 49 years (19-69) with a sex ratio of 3.3. Forty five tumors (86.5%) were classified as stages III - IV according to the 2017 UICC TNM classification. Loco regional relapse (LRR) was defined as a local and/or regional progression that occurs at least 6 months after the end of treatment. Survival analysis was performed according to Kaplan-Meier method and Log-rank test was used to compare anatomy clinical and therapeutic factors that may influence loco regional free survival (LRFS). Results: After a median follow up of 42 months, 6 patients (11.5%) experienced LRR. A metastatic relapse was also noted for 3 of these patients (50%). Target volumes coverage was optimal for all patient with LRR. Four relapses (66.6%) were in high-risk target volume and two (33.3%) were borderline. Three years LRFS was 85,9%. Four factors predicted loco regional relapses: histologic type other than undifferentiated (UCNT) (p=0.027), a macroscopic pre chemotherapy tumor volume exceeding 100 cm³ (p=0.005), a reduction in IC doses exceeding 20% (p=0.016) and a total cumulative cisplatin dose less than 380 mg/m² (p=0.0.34). TNM classification and response to IC did not impact loco regional relapses. Conclusion: For nasopharyngeal carcinoma, tumors with initial high volume and/or histologic type other than UCNT, have a higher risk of loco regional relapse. Therefore, they require a more aggressive therapeutic approaches and a suitable monitoring protocol.

Keywords: loco regional relapse, modulation intensity radiotherapy, nasopharyngeal carcinoma, prognostic factors

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