Search results for: immune organs
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1135

Search results for: immune organs

1015 Effects of a Bacteria-Based Probiotic on Subpopulations of Peripheral Leukocytes and Their Interleukin mRNA Expression in Calves

Authors: Abdul Qadir Qadis, Satoru Goya, Minoru Yatsu, Yu-uki Yoshida, Toshihiro Ichijo, Shigeru Sato

Abstract:

Bacterial probiotics are known to modulate the gut-associated lymphoid and epithelial tissue response to enhance the activities of intestinal and systemic immune system in human and animals. In cattle, the immune-stimulatory effects of probiotics have been evaluated during intestinal disorders. To investigate the effects of probiotic on the function of peripheral blood mononuclear cells, eight healthy Holstein calves (10 ± 3 weeks) were assigned to a 4 × 2 experimental design. The probiotic, consisting of Lactobacillus plantarum, Enterococcus faecium and Clostridium butyricum, was administered orally at 3.0 g/100 kg body weight to calves once daily for 5 consecutive days. Calves given no probiotic served as the control. In the treatment group, increases in numbers of CD282+ monocytes, CD3+ T-cells and CD4+, CD8+ and WC1+ γδ T- cell subsets were noted on day 7 post-placement compared to pre-dose day and the control group. Expression of interleukin-6, interferon-gamma and tumor necrosis factor-alpha was elevated in peripheral leukocytes on days 7 and 14. These results suggest that peripheral blood leukocytes in healthy calves may be stimulated via the gastrointestinal microbiota, which was increased by the oral probiotic treatment. The 5-day repeated administration of a bacterial probiotic may enhance cellular immune function in weaned calves.

Keywords: bacterial-probiotic, calf, interleukin, leukocyte

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1014 The Dose to Organs in Lumbar-Abdominal Computed Tomography Imaging Using TLD

Authors: M. Zehtabian, Z. Molaiemanesh, Z. Shafahi, M. Papie, M. Zahraie Moghaddam, M. Mehralizadeh, M. R. Vahidi, S. Sina

Abstract:

The introduction of CT scans has been a great improvement in diagnosis of different diseases. However, this imaging modality can expose the patients to cumulative radiation doses which may increase the risks of some health problems like cancer. In this study, the dose delivered to different organs in lumbar-abdominal imaging was measured by putting the TLD-100, and TLD-100H chips inside the Alderson Rando phantom. The lumbar-abdominal image of the phantom was obtained, while TLD chips were inside the holes of the phantom. According to the results obtained in this study using TLD-100 chips, the average dose received by liver, bladder, rectum, kidneys, and uterus were found to be 12.9 mSv, 8.9 mSv, 10.1 mSv, 11.0 mSv, 11.2 mSv, and 10.5 mSv respectively, while the measurements performed by TLD-100H show that the average dose to liver, bladder, rectum, kidneys, and uterus were found to be 12.4 mSv, 9.2 mSv, 9.5 mSv, 10.5 mSv, 10.7 mSv, and 9.9 mSv respectively. The results of this study indicates that the dose measured by the TLD-100H chips are in close agreement with those obtained by TLD-100.

Keywords: CT scan, dose, TLD-100, diagnosis

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1013 Kids and COVID-19: They are Winning with Their Immunity

Authors: Husham Bayazed, Fatimah Yousif

Abstract:

Purpose of Presentation: The infant immune system has a reputation for being weak and underdeveloped when compared to the adult immune system, but the comparison isn’t quite fair. At the start, as the COVID-19 pandemic drags on and evolves, many Pediatricians and kids' parents have been left with renewed questions about the consequences and sequel of infection on children and the steps to be taken if their child has the symptoms of COVID-19 or tests positive. Recent Findings Literature reviews and recent studies revealed that children are better than adults at controlling SARS-CoV-2. There was conflicting evidence on age-related differences in ACE2 expression in the nose and lungs. But scientists who measured the ‘viral load’ in children's upper airways have seen no clear difference between children and adults. Moreover, the hypothesis is that kids might be more exposed to other coronaviruses common cold, with a production of ready protective antibodies to lock on to the pandemic coronavirus. But the evidence suggests that adults also have this immunity too. Strikingly, these ‘cross-reactive’ antibodies don’t offer any special protection. Summary One of the few silver linings of the Covid-19 pandemic is that children are relatively spared. The kid's Innate Immunity is hardly the whole story, the innate immune response against SARS-CoV-2 infection is early initiative calm with low immunological tone to prevent an overactive immunity and with rapidly repair damage to the lungs in contrast to stormy waves in adults. Therefore, Kids are at much lower risk of Covid-19 infection, and they are still winning the battle against Covid-19 with their innate immunity.

Keywords: Covid-19, kids, ACE2 receptors, immunity

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1012 Evolution of Mineral Nutrition in Two Species of Atriplex (halimus and canescens) under Salt Stress

Authors: Z. Mahi, L. Marousset, C. Roudaut, M. Belkhodja, R. Lemoine

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The strong accumulation of salts in the soil as well as in irrigation water greatly disrupts the growth and development of almost all plants. The study of these disturbances in halophytes helps provide better guidance on the deteriorating effect of salinity. Evaluation of salt stress in two species of Atriplex (halimus and canescens) through the study of mineral nutrition (dosage of sodium and potassium) shows a variability of responses. The results show that the Na+ ion accumulates in the three organs whatever the applied concentration. This accumulation increases with the high salt concentrations in halimus whereas in canescens, 600 mM treatment shows a reduction of the amount of this element. A decrease in the amount of potassium is observed for all organs except halimus rods 100 mM. Unlike halimus, canescens K + accumulates in high concentrations of salt at the roots and leaves. The ratio Na+/K+ decreases the salt by halimus against it increases in levels canescens roots and treated with high concentrations of NaCl (600 mM) leaves.

Keywords: Atriplex, canescens, halimus, Na +, K +, Na Cl, tolerance

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1011 Study the Action of Malathion Induced Enzymatic Changes in the Target Organ of Fish Labeo Rohita

Authors: Sudha Summarwar, Jyotsana Pandey, Deepali Lall

Abstract:

The Malathion compound has the great tendency to be accumulated in the organs of the fishes both if it is present in traces or in higher amount in the aquatic environment. It has the tendency to be accumulated more in quantity in the organs directly exposed to it. The accumulation was found to be time and concentration dependent. The accumulation of malathion was maximum in gills and is the minimum in the brain. Effect of different sub-lethal concentrations (l/5th, l/l0th, l/15th, l/20th, and 1/25th fractions of 96 hr. LC50) of malathion compound on acid phosphatase (AcPase), alkaline phosphatase (AlPase), serum glutamic oxalacetic transaminase (SGOT) and Serum Glucose-6-Phosphatase (S-G-6-Pase), serum glutamic pyruvic transaminase (SGPT) in blood of Labeo rohita exposed for the period of 15. 30, 45, and 60 days, have been studied in present investigations. In general the alterations were concentrations and duration dependent.

Keywords: AcPase, AlPase, Labeo rohita, malathion, S-G-6-Pase, SGOT, SGPT

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1010 Introduce a New Model of Anomaly Detection in Computer Networks Using Artificial Immune Systems

Authors: Mehrshad Khosraviani, Faramarz Abbaspour Leyl Abadi

Abstract:

The fundamental component of the computer network of modern information society will be considered. These networks are connected to the network of the internet generally. Due to the fact that the primary purpose of the Internet is not designed for, in recent decades, none of these networks in many of the attacks has been very important. Today, for the provision of security, different security tools and systems, including intrusion detection systems are used in the network. A common diagnosis system based on artificial immunity, the designer, the Adhasaz Foundation has been evaluated. The idea of using artificial safety methods in the diagnosis of abnormalities in computer networks it has been stimulated in the direction of their specificity, there are safety systems are similar to the common needs of m, that is non-diagnostic. For example, such methods can be used to detect any abnormalities, a variety of attacks, being memory, learning ability, and Khodtnzimi method of artificial immune algorithm pointed out. Diagnosis of the common system of education offered in this paper using only the normal samples is required for network and any additional data about the type of attacks is not. In the proposed system of positive selection and negative selection processes, selection of samples to create a distinction between the colony of normal attack is used. Copa real data collection on the evaluation of ij indicates the proposed system in the false alarm rate is often low compared to other ir methods and the detection rate is in the variations.

Keywords: artificial immune system, abnormality detection, intrusion detection, computer networks

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1009 Analysis of the Presence of Alkylglycerols by Gas Chromatography in Ostrich Oil

Authors: Luana N. Cardozo, Debora A. S. Coutinho, Fabiola Lagher, Bruno J. G. Silva, Ivonilce Venture, Mainara Tesser, Graciela Venera

Abstract:

Ostrich oil is used as food in Brazil, and it has been the subject of scientific research because it contains essential fatty acids (Omega 3, 6, 7, and 9), which provide benefits to human health. Alkylglycerols are lipid ethers consisted of a saturated or unsaturated hydrocarbon chain joined by ether-type bonding to one of the glycerol hydroxyls. It is known that supplementation with alkylglycerols can act significantly on the functioning of immune system cells, both in pathological situations and in homeostasis. Objective: Analyze the presence of alkylglycerols in ostrich oil. Methods: The ostrich oil was bought from an industry that manufactures the product for sale as food, located in Mirante da Serra, northern Brazil. The samples were sent for analysis to the chemistry department of the Federal University of Paraná, where they were analyzed by the gas chromatography method. Results: The analysis of the ostrich oil presented alkylglycerols in area 514505154. Comparison, it is possible to observe that shark liver oil contains the area 26190196, and the difference between both is highly significant. Conclusion: The importance of alkylglycerol supplementation for the immune system is known. The analysis of the results made it possible to verify the presence of alkylglycerols in the ostrich oil, which is five times higher than in the shark liver oil, that would be the largest source food, but was surpassed by the ostrich oil until the present time. The present study emphasizes that ostrich oil can be considered a food source of alkylglycerols and may play a promising role in the immune system because it contains such substance, but further studies are needed to prove its performance in the body.

Keywords: ostrich oil, nutritional composition, alkylglycerols, food

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1008 Targeted Photodynamic Therapy for Intraperitoneal Ovarian Cancer, A Way to Stimulate Anti-Tumoral Immune Response

Authors: Lea Boidin, Martha Baydoun, Bertrand Leroux, Olivier Morales, Samir Acherar, Celine Frochot, Nadira Delhem

Abstract:

Ovarian cancer (OC) is one of the most defying diseases in gynecologic oncology. Even though surgery remains crucial in the therapy of patients with primary ovarian cancer, recurrent recidivism calls for the development of new therapy protocols to propose for patients dealing with this cancer. FRα is described as a tumor‐associated antigen in OC, where FRα expression is usually linked with more poorly differentiated, aggressive tumors. The Photodynamic treatment (PDT) available data have shown improvements in the uptake of small tumors and in the induction of a proper anti-tumoral immune response. In order to target specifically peritoneal metastatis, which overexpress FRα, a new-patented PS coupled with folic acid has been developed in our team. Herein we propose PDT using this new patented PS for PDT applied in an in vivo mice model. The efficacy of the treatment was evaluated in mice without and with PBMC reconstitution. Mice were divided into four groups: Non-Treated, PS, Light Only, and PDT Treated and subjected to illumination by laser set at 668nm with a duration of illumination of 45 minutes (or 1 min of illumination followed by 2 minutes of pause repeated 45 times). When mice were not reconstituted and after fractionized PDT protocol, a significant decrease in the tumor volume was noticed. An induction in the anti-tumoral cytokine IFNγ chaperoned this decrease while a subsequent inhibition in the cytokine TGFβ. Even more crucial, when mice were reconstituted and upon PDT, the fold of tumor decrease was even higher. An immune response was activated decoded with an increase in NK, CD3 +, LT helper and Cytotoxic T cells. Thereafter, an increase in the expression of the cytokines IFNγ and TNFα were noticed while an inhibition in TGFβ, IL8 and IL10 accompanied this immune response activation. Therefore, our work has shown for the first time that a fractionized PDT protocol using a folate-targeted PDT is effective for treatment of ovarian cancer. The interest in using PDT in this case, goes beyond the local induction of tumor apoptosis only, but can promote subsequent anti-tumor response. Most of the therapies currently used to treat ovarian cancer, have an uncooperative outcomes on the host immune response. The readiness of a tumor adjuvant treatment like PDT adequate in eliminating the tumor and in concert stimulating anti-tumor immunity would be weighty.

Keywords: folate receptor, ovarian cancer, photodynamic therapy, humanized mice model

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1007 Autoantibodies against Central Nervous System Antigens and the Serum Levels of IL-32 in Patients with Schizophrenia

Authors: Fatemeh Keshavarz

Abstract:

Background: Schizophrenia is a disease of the nervous system, and immune system disorders can affect its pathogenesis. Activation of microglia, proinflammatory cytokines, disruption of the blood-brain barrier (BBB) due to inflammation, activation of autoreactive B cells, and consequently the production of autoantibodies against system antigens are among the immune processes involved in neurological diseases. interleukin 32 (IL-32) a proinflammatory cytokine that important player in the activation of the innate and adaptive immune responses. This study aimed to measure the serum level of IL-32 as well as the frequency of autoantibody positivity against several nervous system antigens in patients with schizophrenia. Material and Methods: This study was conducted on 40 patients with schizophrenia and 40 healthy individuals in the control group. Serum IL-32 levels were measured by ELISA. The frequency of autoantibodies against Hu, Ri, Yo, Tr, CV2, Amphiphysin, SOX1, Zic4, ITPR1, CARP, GAD, Recoverin, Titin, and Ganglioside antigens were measured by indirect immunofluorescence method. Results: Serum IL-32 levels in patients with schizophrenia were significantly higher compared to the control group. Autoantibodies were positive in 8 patients for GAD antigen and 5 patients for Ri antigen, which showed a significant relationship compared to the control group. Autoantibodies were also positive in 2 patients for CV2, in 1 patient for Hu, and in 1 patient for CARP. Negative results were reported for other antigens. Conclusion: Our findings suggest that elevated the serum IL-32 level and autoantibody positivity against several nervous system antigens may be involved in the pathogenesis of schizophrenia.

Keywords: schizophrenia, microglia, autoantibodies, IL-32

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1006 Effect of Peganum harmala Seeds on Blood Factors, Immune Response and Intestinal Selected Bacterial Population in Broiler Chickens

Authors: Majid Goudarzi

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This experiment was designed to study the effects of feeding different levels of Peganum harmala seeds (PHS) and antibiotic on serum biochemical parameters, immune response and intestinal microflora composition in Ross broiler chickens. A total of 240 one-d-old unsexed broiler chickens were randomly allocated to each of the four treatment groups, each with four replicate pens of 15 chicks. The dietary treatments included of control (C) - without PHS and antibiotic - the diet contains 300 mg/kg Lincomycin 0.88% (A) and the diets contain 2 g/kg (H1) and 4 g/kg (H2) PHS. The chicks were raised on floor pens and received diets and water ad libitum for six weeks. Blood samplings were performed for the determination of antibody titer against Newcastle disease on 14 and 21 days and for biochemical parameters on 42 days of age. The populations of Lactobacilli spp. and Escherichia coli were enumerated in ileum by conventional microbiological techniques using selective agar media. Inclusion of PHS in diet resulted in a significant decrease in total cholesterol and significant increase in HDL relative to the control and antibiotic groups. Antibody titer against NDV was not affected by experimental treatments. E. coli population in birds supplemented with antibiotic and PHS was significantly lower than control, but Lactobacilli spp. population increased only by antibiotic and not by PHS. In conclusion, the results of this study showed that addition of PHS powder seem to have a positive influence on some biochemical parameters and gastrointestinal microflora.

Keywords: antibiotic, biochemical parameters, immune system, Peganum harmala

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1005 Absorbed Dose Estimation of 177Lu-DOTATOC in Adenocarcinoma Breast Cancer Bearing Mice

Authors: S. Zolghadri, M. Mousavi-Daramoroudi, H. Yousefnia, F. Abbasi-Davani

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In this study, the absorbed dose of human organs after injection of 177Lu-DOTATOC was studied based on the biodistribution of the complex in adenocarcinoma breast cancer bearing mice. For this purpose, the biodistribution of the radiolabelled complex was studied and compartmental modeling was applied to calculate the absorbed dose with high precision. As expected, 177Lu-DOTATOC illustrated a notable specific uptake in tumor and pancreas, organs with high level of somatostatin receptor on their surface and the effectiveness of the radio-conjugate for targeting of the breast adenocarcinoma tumors was indicated. The elicited results of modeling were the exponential equations, and those are utilized for obtaining the cumulated activity data by taking their integral. The results also exemplified that non-target absorbed-doses such as the liver, spleen and pancreas were approximately 0.008, 0.004, and 0.039, respectively. While these values were so much lower than target (tumor) absorbed-dose, it seems due to this low toxicity, this complex is a good agent for therapy.

Keywords: ¹⁷⁷Lu, breast cancer, compartmental modeling, dosimetry

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1004 Effector and Memory Immune Responses Induced by Total Extracts of Naegleria fowleri Co-Administered with Cholera Toxin

Authors: Q. B. Maria de la Luz Ortega Juárez, Saúl Rojas Hernández, Itzel Berenice Rodríguez Mera, María Maricela Carrasco Yépez, Mara Gutierrez Sánchez

Abstract:

Naegleria fowleri is a free-living amoeba found mainly in temperate freshwater and is the etiologic agent of primary amebic meningoencephalitis (PAM), a fatal acute disease with a mortality rate greater than 95%. At present, there are no treatments available for MAP, and the development of effective vaccines that generate long-term immunological memory allowing protection against MAP would be of great importance. The objective of this work was to analyze the effector and memory immune response in BALB/c mice immunized with total extract of N. fowleri co-administered with cholera toxin. In this study, BALB/c mice were immunized four times intranasally with ET of N. fowleri adjuvanted with CT with or without booster at three months and were challenged or not with the lethal dose of N. fowleri, determining survival, the humoral, effector and memory response, by ELISA and flow cytometry techniques. The results obtained showed that the survival of mice immunized with booster had 60% protection compared to the group without booster, which obtained 20% protection. Evaluating the humoral response, it was found that both IgG and IgA levels were higher in sera than in nasal washes in both treatments. In the cellular response, the increase in the percentage of positive cells was found for effector T and B lymphocytes in the nasal passages (NP) in the group with boost and nasopharynx-associated lymphoid tissue (NALT) in the group without boost and lymphocytes only. B in both treatments, as well as in memory cells treatment with boost T lymphocytes in PN and NALT and without boost in cervical lymph nodes (CG) with respect to B lymphocytes, in PN, GC and NALT in treatment with boost and NALT in treatment without booster. Therefore, the involvement of the effector immune response and memory play a fundamental role for protection against N. fowleri and for the development of vaccine candidates.

Keywords: immune response, immunological memory, naegleria fowleri, primary amebic meningoencephalitis

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1003 Nutrition Bio-Shield Superfood: Healthy and Live Herbal Supplement for Immune System Enhancement

Authors: Azam Bayat, Aref Khalkhali, Ali Reza Mahjoub

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Healthy and viable herbal supplement were prepared from wheat by a green route. This organic biomaterial was named Nutrition Bio-shield Superfood (NBS). The NBS supplement had various vitamins, macro and micro molecules, and ingredients. In this study, 20 small Balb/C labile specimens were used in a weighing 30 ± 5 range. The samples were randomly divided into different groups, then the groups were divided into 5 groups. According to the results of this study, the mean number of white blood cells and neutrophil percentage in the experimental group receiving healthy and live dietary supplement showed a significant increase at the 5% probability level in all three groups received 50, 100 and 150 mg/ kg body weight of the mouse compared to the control group. In general, the dietary supplement increases the level of immunity.

Keywords: healthy and live herbal supplement, biomaterial, immune system, enhancement

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1002 Design of an Improved Distributed Framework for Intrusion Detection System Based on Artificial Immune System and Neural Network

Authors: Yulin Rao, Zhixuan Li, Burra Venkata Durga Kumar

Abstract:

Intrusion detection refers to monitoring the actions of internal and external intruders on the system and detecting the behaviours that violate security policies in real-time. In intrusion detection, there has been much discussion about the application of neural network technology and artificial immune system (AIS). However, many solutions use static methods (signature-based and stateful protocol analysis) or centralized intrusion detection systems (CIDS), which are unsuitable for real-time intrusion detection systems that need to process large amounts of data and detect unknown intrusions. This article proposes a framework for a distributed intrusion detection system (DIDS) with multi-agents based on the concept of AIS and neural network technology to detect anomalies and intrusions. In this framework, multiple agents are assigned to each host and work together, improving the system's detection efficiency and robustness. The trainer agent in the central server of the framework uses the artificial neural network (ANN) rather than the negative selection algorithm of AIS to generate mature detectors. Mature detectors can distinguish between self-files and non-self-files after learning. Our analyzer agents use genetic algorithms to generate memory cell detectors. This kind of detector will effectively reduce false positive and false negative errors and act quickly on known intrusions.

Keywords: artificial immune system, distributed artificial intelligence, multi-agent, intrusion detection system, neural network

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1001 Interaction between Kazal-Type Serine Proteinase Inhibitor SPIPm2 and Cyclophilin A from the Black Tiger Shrimp Penaeus monodon

Authors: Sirikwan Ponprateep, Anchalee Tassanakajon, Vichien Rimphanitchayakit

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A Kazal-type serine proteinase inhibitor, SPIPm2, was abundantly expressed in the hemocytes and secreted into shrimp plasma has anti-viral property against white spot syndrome virus (WSSV). To discover the molecular mechanism of antiviral activity, the binding assay showed that SPIPm2 bind to the components of viral particle and shrimp hemocyte. From our previous report, viral target protein of SPIPm2 was identified, namely WSV477 using yeast two-hybrid screening. WSV477 is an early gene product of WSSV and involved in viral propagation. In this study, the co-immunoprecipitation technique and Tandem Mass Spectrometry (LC-MS/MS) was used to identify the target protein of SPIPm2 from shrimp hemocyte. The target protein of SPIPm2 was cyclophilin A. In vertebrate, cyclophilin A or peptidylprolyl isomerase A was reported to be the immune suppressor interacted with cyclosporin A involved in immune defense response. The recombinant cyclophilin A from Penaeus monodon (rPmCypA) was produced in E.coli system and purified using Ni-NTA column to confirm the protein-protein interaction. In vitro pull-down assay showed the interaction between rSPIPm2 and rPmCypA. To study the biological function of these proteins, the expression analysis of immune gene in shrimp defense pathways will be investigated after rPmCypA administration.

Keywords: cyclophilin A, protein-protein interaction, Kazal-type serine proteinase inhibitor, Penaeus monodon

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1000 Poly(Amidoamine) Dendrimer-Cisplatin Nanocomplex Mixed with Multifunctional Ovalbumin Coated Iron Oxide Nanoparticles for Immuno-Chemotherapeutics with M1 Polarization of Macrophages

Authors: Tefera Worku Mekonnen, Hiseh Chih Tsai

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Enhancement of drug efficacy is essential in cancer treatment. The immune stimulator ovalbumin (Ova)-coated citric acid (AC-)-stabilized iron oxide nanoparticles (AC-IO-Ova NPs) and enhanced permeability and retention (EPR) based tumor targeted 4.5 (4.5G) poly(amidoamine) dendrimer-cisplatin nanocomplex (4.5GDP-Cis-pt NC) were used for enhanced anticancer efficiency. The formations of 4.5GDP-Cis-pt NC, AC-IO, and AC-IO-Ova NPs have been examined by FTIR, X-ray diffraction, Raman, and X-ray photoelectron spectroscopy. The conjugation of cisplatin (Cis-pt) with 4.5GDP was confirmed using carbon NMR. The tumor-specific 4.5GDP-Cis-pt NC provided ~45% and 28% cumulative cisplatin release in 72 h at pH 6.5 and 7.4, respectively. A significant immune response with high TNF-α and IL-6 cytokine secretion was confirmed when the co-incubation of AC-IO-Ova with RAW 264.7 or HaCaT cells. AC-IO-Ova NP was biocompatible in different cell lines, even at a high concentration (200 µg mL−1). In contrast, AC-IO-Ova NPs mixed with 4.5GDP-Cis-pt NC (Cis-pt at 15 µg mL−1) significantly increased the cytotoxicity against the cancer cells, which is dose-dependent on the concentration of AC-IO-Ova NPs. The increased anticancer effects may be attributed to the generation of reactive oxygen species (ROS). Moreover, the efficiency of anticancer cells may be further assisted by induction of an innate immune response via M1 macrophage polarization due to the presence of AC-IO-Ova NPs. We provide a better synergestic chemoimmunotherapeutic strategy to enhance the efficiency of anticancer of cisplatin via chemotherapeutic agent 4.5GDP-Cis-pt NC and induction of proinflammatory cytokines to stimulate innate immunity through AC-IO-Ova NPs against tumors.

Keywords: cisplatin-release, iron oxide, ovalbumin, poly(amidoamine) dendrimer

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999 Performance and Physiological Responses of Broiler Chickens to Diets Supplemented with Propolis in Breeding, to in Ovo Propolis Feeding or to Propolis Supplementation of Diets for Their Chicks

Authors: Kalbiye Konanc, Ergin Ozturk

Abstract:

To examine the effects of an ethanol liquid extract obtained from raw bee propolis (PE) on fattening performance and physiology such as vaccine-antibody relationship, microbial profile, immune status and some blood parameters of broiler chickens were used a total of 600 broiler (Ross 308) chicks, obtained from eggs of 288, 38-weeks-old broiler breeding. There were 6 groups: CC (Parent-Control and Offspring-Control, CP (Parent-Control and Offspring-propolis extract, Cip (Parent-Control and Offspring-in-ovo propolis extract), Cis (Parent-Control and Chickens-in-ovo saline), PeC (Parent-propolis extract and Offspring-Control), PeP (Parent-Propolis extract and Offspring-Propolis extract). Each group was consisted of 10 replications with 10 broiler offspring, and the experiment was lasted for 6 weeks with ethanol-extracted propolis concentration is 400 ppm/kg diet. While the highest feed consumptions at 0-21 days and 0-42 days were found in PeC, the best feed conversion ratio at 0-42 days was found in CP group. The live weight gains were found not to be different among the groups. The highest alanine aminotransferase activities were found in CC and CP and aspartate aminotransferase activities in PeP and PeC groups. The highest triglyceride and total antioxidant levels were found highest in CC and the highest total oxidant level in Cip group. IgA level in hatched eggs and IgM value after slaughtering were highest in Cip group. The best immune response was obtained for 21st day Newcastle Disease vaccine in CC and Cis groups and for 28th day Infectious Bursal Disease vaccine in CP group. The highest total aerobic microorganism and the lowest total fungi count were found in PeP group. In conclusion, it was determined that in-ovo propolis ethanol extract (Cip) increased the maternal antibody levels, that had not consistent effects on blood biochemical parameters except for triglyceride, that led to decrease in E. coli counts and that it can provide strong immune response against Infectious Bursal Disease.

Keywords: bee propolis, in-ovo feeding, immune parameters, poultry, maternal antibody, microorganisms

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998 Protection and Immune Responses of DNA Vaccines Targeting Virulence Factors of Streptococcus iniae in Nile Tilapia (Oreochromis niloticus)

Authors: Pattanapon Kayansamruaj, Ha Thanh Dong, Nopadon Pirarat, Channarong Rodkhum

Abstract:

Streptococcus iniae (SI) is a devastating pathogenic bacteria causing heavy mortality in farmed fish. The application of commercialized bacterin vaccine has been reported failures as the outbreaks of the new serotype of SI were emerged in farms after vaccination and subsequently caused severe losses. In the present study, we attempted to develop effective DNA vaccines against SI infection using Nile tilapia (Oreochromis niloticus) as an animal model. Two monovalent DNA vaccines were constructed by the insertion of coding sequences of cell wall-associated virulence factors-encoding genes, comprised of eno (α-enolase) and mtsB (hydrophobic membrane protein), into cytomegalovirus expression vector (pCI-neo). In the animal trial, 30-g Nile tilapia were injected intramuscularly with 15 µg of each vaccine (mock vaccine group was injected by naked pCI-neo) and maintained for 35 days prior challenging with pathogenic SI at the dosage of 107 CFU/fish. At 13 days post-challenge, the relative percent survival of pEno, pMtsB and mock vaccine were 57%, 45% and 27%, respectively. The expression levels of immune responses-associated genes, namely, IL1β, TNF-α, TGF-β, COX2, IL-6, IL-12 and IL-13, were investigated from the spleen of experimental animal at 7 days post-vaccination (PV) and 7 days post-challenge (PC) using quantitative RT-PCR technique. Generally, at 7 days PV, the pEno vaccinated group exhibited highest level of up-regulation (1.7 to 2.9 folds) of every gene, but TGF-β, comparing to pMtsB and mock vaccine groups. However, at 7 days PC, pEno group showed significant up-regulation (1.4 to 8.5 folds) of immune-related genes as similar as mock vaccine group, while pMtsB group had lowest level of up-regulation (0.7 to 3.3 folds). Summarily, this study indicated that the pEno and pMtsB vaccines could elicit the immune responses of the fish and the magnitude of gene expression at 7 days PV was also consistent with the protection level conferred by the vaccine.

Keywords: gene expression, DNA vaccine, Nile tilapia, Streptococcus iniae

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997 Determination of Critical Organ Doses for Liver Scintigraphy Using Cr-51

Authors: O. Maranci, A. B. Tugrul

Abstract:

Scintigraphy is an imaging method of nuclear events provoked by collisions or charged current interactions with radiation. It is used for diagnostic test used in nuclear medicine via radiopharmaceuticals emitting radiation which is captured by gamma cameras to form two-dimensional images. Liver scintigraphy is widely used in nuclear medicine.Tc-99m and Cr-51 gamma radioisotopes can be used for this purpose. Cr-51 usage is more important for patients’ organ dose that has higher energy and longer half-life as compared to Tc-99m. In this study, it is aimed to determine the required dose for critical organs of patient through liver scintigraphy via Cr-51 gamma radioisotope. Experimental studies were conducted on patients even though conducting experimental studies on patients is extremely difficult for determination of critical organ doses. Torso phantom was utilized to simulate the liver scintigraphy by using 20 mini packages of Cr-51 that were placed on the organ. The radioisotope was produced by irradiation in central thimble of TRIGA MARK II Reactor at 250 KW power. As the results of the study, critical organ doses were determined and evaluated with different critic organs.

Keywords: critical organ doses, liver, scintigraphy, TRIGA Mark-II

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996 Effect of Lowering the Proportion of Chlorella vulgaris in Fish Feed on Tilapia's Immune System

Authors: Hamza A. Pantami, Khozizah Shaari, Intan S. Ismail, Chong C. Min

Abstract:

Introduction: Tilapia is the second-highest harvested freshwater fish species in Malaysia, available in almost all fish farms and markets. Unfortunately, tilapia culture in Malaysia is highly affected by Aeromonas hydrophila and Streptococcus agalactiae, which affect the production rate and consequently pose a direct negative economic impact. Reliance on drugs to control or reduce bacterial infections has been led to contamination of water bodies and development of drug resistance, as well as gave rise to toxicity issues in downstream fish products. Resorting to vaccines have helped curb the problem to a certain extent, but a more effective solution is still required. Using microalgae-based feed to enhance the fish immunity against bacterial infection offers a promising alternative. Objectives: This study aims to evaluate the efficacy of Chlorella vulgaris at lower percentage incorporation in feeds for an immune boost of tilapia in a shorter time. Methods: The study was in two phases. The safety concentration studies at 500 mg/kg-1 and the administration of cultured C. vulgaris biomass via incorporation into fish feed for five different groups in three weeks. Group 1 was the control (0% incorporation), whereas group 2, 3, 4 and 5 received 0.625%, 1.25%, 2.5% and 5% incorporation respectively. The parameters evaluated were the blood profile, serum lysozyme activity (SLA), serum bactericidal activity (SBA), phagocytosis activity (PA), respiratory burst activity (RBA), and lymphoproliferation activity (LPA). The data were analyzed via ANOVA using SPSS (version 16). Further testing was done using Tukey’s test. All tests were performed at the 95% confidence interval (p < 0.05). Results: There were no toxic signs in tilapia fish at 500 mg/kg-1. Treated groups showed significantly better immune parameters compared to the control group (p < 0.05). Conclusions: C. vulgaris crude biomass in a fish meal at a lower incorporation level of 5% can increase specific and non-specific immunity in tilapia fish in a shorter time duration.

Keywords: Chlorella vulgaris, hematology profile, immune boost, lymphoproliferation

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995 Pretherapy Initial Dosimetry Results in Prostat Cancer Radionuclide Therapy with Lu-177-PSMA-DOTA-617

Authors: M. Abuqebitah, H. Tanyildizi, N. Yeyin, I. Cavdar, M. Demir, L. Kabasakal

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Aim: Targeted radionuclide therapy (TRT) is an increasingly used treatment modality for wide range of cancers. Presently dosimetry is highly required either to plan treatment or to ascertain the absorbed dose delivered to critical organs during treatment. Methods and Materials: The study comprised 7 patients suffered from prostate cancer with progressive disease and candidate to undergo Lu-177-DOTA-617 therapy following to PSMA- PET/CT imaging for all patients. (5.2±0.3 mCi) was intravenously injected. To evaluate bone marrow absorbed dose 2 cc blood samples were withdrawn in short variable times (3, 15, 30, 60, 180 minutes) after injection. Furthermore, whole body scans were performed using scintillation gama camera in 4, 24, 48, and 120 hours after injection and in order to quantify the activity taken up in the body, kidneys , liver, right parotid, and left parotid the geometric mean of anterior and posterior counts were determined through ROI analysis, after that background subtraction and attenuation correction were applied using patients PSMA- PET/CT images taking in a consideration: organ thickness, body thickness, and Hounsfield unites from CT scan. OLINDA/EXM dosimetry program was used for curve fitting, residence time calculation, and absorbed dose calculations. Findings: Absorbed doses of bone marrow, left kidney, right kidney, liver, left parotid, right parotid, total body were 1.28±0.52, 32.36±16.36, 32.7±13.68, 10.35±3.45, 38.67±21.29, 37.55±19.77, 2.25±0.95 (mGy/mCi), respectively. Conclusion: Our first results clarify that Lu-177-DOTA-617 is safe and reliable therapy as there were no complications seen. In the other hand, the observable variation in the absorbed dose of the critical organs among the patients necessitate patient-specific dosimetry approach to save body organs and particularly highly exposed kidneys and parotid gland.

Keywords: Lu-177-PSMA, prostate cancer, radionuclide therapy

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994 Calculation of Organ Dose for Adult and Pediatric Patients Undergoing Computed Tomography Examinations: A Software Comparison

Authors: Aya Al Masri, Naima Oubenali, Safoin Aktaou, Thibault Julien, Malorie Martin, Fouad Maaloul

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Introduction: The increased number of performed 'Computed Tomography (CT)' examinations raise public concerns regarding associated stochastic risk to patients. In its Publication 102, the ‘International Commission on Radiological Protection (ICRP)’ emphasized the importance of managing patient dose, particularly from repeated or multiple examinations. We developed a Dose Archiving and Communication System that gives multiple dose indexes (organ dose, effective dose, and skin-dose mapping) for patients undergoing radiological imaging exams. The aim of this study is to compare the organ dose values given by our software for patients undergoing CT exams with those of another software named "VirtualDose". Materials and methods: Our software uses Monte Carlo simulations to calculate organ doses for patients undergoing computed tomography examinations. The general calculation principle consists to simulate: (1) the scanner machine with all its technical specifications and associated irradiation cases (kVp, field collimation, mAs, pitch ...) (2) detailed geometric and compositional information of dozens of well identified organs of computational hybrid phantoms that contain the necessary anatomical data. The mass as well as the elemental composition of the tissues and organs that constitute our phantoms correspond to the recommendations of the international organizations (namely the ICRP and the ICRU). Their body dimensions correspond to reference data developed in the United States. Simulated data was verified by clinical measurement. To perform the comparison, 270 adult patients and 150 pediatric patients were used, whose data corresponds to exams carried out in France hospital centers. The comparison dataset of adult patients includes adult males and females for three different scanner machines and three different acquisition protocols (Head, Chest, and Chest-Abdomen-Pelvis). The comparison sample of pediatric patients includes the exams of thirty patients for each of the following age groups: new born, 1-2 years, 3-7 years, 8-12 years, and 13-16 years. The comparison for pediatric patients were performed on the “Head” protocol. The percentage of the dose difference were calculated for organs receiving a significant dose according to the acquisition protocol (80% of the maximal dose). Results: Adult patients: for organs that are completely covered by the scan range, the maximum percentage of dose difference between the two software is 27 %. However, there are three organs situated at the edges of the scan range that show a slightly higher dose difference. Pediatric patients: the percentage of dose difference between the two software does not exceed 30%. These dose differences may be due to the use of two different generations of hybrid phantoms by the two software. Conclusion: This study shows that our software provides a reliable dosimetric information for patients undergoing Computed Tomography exams.

Keywords: adult and pediatric patients, computed tomography, organ dose calculation, software comparison

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993 Effect of Different Levels of Distillery Yeast Sludge on Immune Level, Egg Quality and Performance of Layers as a Substitute for Soybean Meal

Authors: Rana Bilal, Faiz-Ul-Hassan, Moazzam Jameel

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There is a dire need to replace high-cost protein with more economical protein to overcome animal protein shortage in developing nations especially countries like Pakistan. In conjunction with these efforts, the current study was planned to evaluate the effects of various dried distillery yeast sludge (DYS) levels on the immune level, egg quality, and performance of layers by replacing soybean meal. The study was designed with two hundred layers of Hy-Line variety. Distillery yeast sludge was dried and ground for 2 mm mesh size and after this proximate and mineral analysis was determined. Five isocaloric and isonitrogeneous feeds were given containing C (control), 5, 10, 15, 20% distillery yeast sludge by replacing soybean meal. The trial was performed in the completely randomized design with five treatments, 4 replicates and 10 hen per replicate. Results demonstrated that feed intake, egg production, feed conversion ratio decreased (P < 0.05) with the increased dietary DYS. However, statistically significant decrease (P < 0.05) was found in hens having DYS20 diet than control. Layers on Diets C, DYS5 and DYS10 exerted a higher immune level than DYS15 and DYS20 diets. Egg weight, eggshell weight, eggshell thickness, egg albumen height as well as haugh unit score were affected significantly by the increased level of DYS. In general, results of this study demonstrated that inclusion of DYS up to 10% showed no adverse effects on health and performance of layers.

Keywords: egg quality, immunity, layers, performance

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992 Chemical vs Visual Perception in Food Choice Ability of Octopus vulgaris (Cuvier, 1797)

Authors: Al Sayed Al Soudy, Valeria Maselli, Gianluca Polese, Anna Di Cosmo

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Cephalopods are considered as a model organism with a rich behavioral repertoire. Sophisticated behaviors were widely studied and described in different species such as Octopus vulgaris, who has evolved the largest and more complex nervous system among invertebrates. In O. vulgaris, cognitive abilities in problem-solving tasks and learning abilities are associated with long-term memory and spatial memory, mediated by highly developed sensory organs. They are equipped with sophisticated eyes, able to discriminate colors even with a single photoreceptor type, vestibular system, ‘lateral line analogue’, primitive ‘hearing’ system and olfactory organs. They can recognize chemical cues either through direct contact with odors sources using suckers or by distance through the olfactory organs. Cephalopods are able to detect widespread waterborne molecules by the olfactory organs. However, many volatile odorant molecules are insoluble or have a very low solubility in water, and must be perceived by direct contact. O. vulgaris, equipped with many chemosensory neurons located in their suckers, exhibits a peculiar behavior that can be provocatively described as 'smell by touch'. The aim of this study is to establish the priority given to chemical vs. visual perception in food choice. Materials and methods: Three different types of food (anchovies, clams, and mussels) were used, and all sessions were recorded with a digital camera. During the acclimatization period, Octopuses were exposed to the three types of food to test their natural food preferences. Later, to verify if food preference is maintained, food was provided in transparent screw-jars with pierced lids to allow both visual and chemical recognition of the food inside. Subsequently, we tested alternatively octopuses with food in sealed transparent screw-jars and food in blind screw-jars with pierced lids. As a control, we used blind sealed jars with the same lid color to verify a random choice among food types. Results and discussion: During the acclimatization period, O. vulgaris shows a higher preference for anchovies (60%) followed by clams (30%), then mussels (10%). After acclimatization, using the transparent and pierced screw jars octopus’s food choices resulted in 50-50 between anchovies and clams, avoiding mussels. Later, guided by just visual sense, with transparent but not pierced jars, their food preferences resulted in 100% anchovies. With pierced but not transparent jars their food preference resulted in 100% anchovies as first food choice, the clams as a second food choice result (33.3%). With no possibility to select food, neither by vision nor by chemoreception, the results were 20% anchovies, 20% clams, and 60% mussels. We conclude that O. vulgaris uses both chemical and visual senses in an integrative way in food choice, but if we exclude one of them, it appears clear that its food preference relies on chemical sense more than on visual perception.

Keywords: food choice, Octopus vulgaris, olfaction, sensory organs, visual sense

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991 A Galectin from Rock Bream Oplegnathus fasciatus: Molecular Characterization and Immunological Properties

Authors: W. S. Thulasitha, N. Umasuthan, G. I. Godahewa, Jehee Lee

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In fish, innate immune defense is the first immune response against microbial pathogens which consists of several antimicrobial components. Galectins are one of the carbohydrate binding lectins that have the ability to identify pathogen by recognition of pathogen associated molecular patterns. Galectins play a vital role in the regulation of innate and adaptive immune responses. Rock bream Oplegnathus fasciatus is one of the most important cultured species in Korea and Japan. Considering the losses due to microbial pathogens, present study was carried out to understand the molecular and functional characteristics of a galectin in normal and pathogenic conditions, which could help to establish an understanding about immunological components of rock bream. Complete cDNA of rock bream galectin like protein B (rbGal like B) was identified from the cDNA library, and the in silico analysis was carried out using bioinformatic tools. Genomic structure was derived from the BAC library by sequencing a specific clone and using Spidey. Full length of rbGal like B (contig14775) cDNA containing 517 nucleotides was identified from the cDNA library which comprised of 435 bp in the open reading frame encoding a deduced protein composed of 145 amino acids. The molecular mass of putative protein was predicted as 16.14 kDa with an isoelectric point of 8.55. A characteristic conserved galactose binding domain was located from 12 to 145 amino acids. Genomic structure of rbGal like B consisted of 4 exons and 3 introns. Moreover, pairwise alignment showed that rock bream rbGal like B shares highest similarity (95.9 %) and identity (91 %) with Takifugu rubripes galectin related protein B like and lowest similarity (55.5 %) and identity (32.4 %) with Homo sapiens. Multiple sequence alignment demonstrated that the galectin related protein B was conserved among vertebrates. A phylogenetic analysis revealed that rbGal like B protein clustered together with other fish homologs in fish clade. It showed closer evolutionary link with Takifugu rubripes. Tissue distribution and expression patterns of rbGal like B upon immune challenges were performed using qRT-PCR assays. Among all tested tissues, level of rbGal like B expression was significantly high in gill tissue followed by kidney, intestine, heart and spleen. Upon immune challenges, it showed an up-regulated pattern of expression with Edwardsiella tarda, rock bream irido virus and poly I:C up to 6 h post injection and up to 24 h with LPS. However, In the presence of Streptococcus iniae rbGal like B showed an up and down pattern of expression with the peak at 6 - 12 h. Results from the present study revealed the phylogenetic position and role of rbGal like B in response to microbial infection in rock bream.

Keywords: galectin like protein B, immune response, Oplegnathus fasciatus, molecular characterization

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990 Development of a Diagnostic Device to Predict Clinically Significant Inflammation Associated with Cardiac Surgery

Authors: Mohamed Majrashi, Patricia Connolly, Terry Gourlay

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Cardiopulmonary bypass is known to cause inflammatory response during open heart surgery. It includes the initiation of different cascades such as coagulation, complement system and cytokines. Although the immune system is body’s key defense mechanism against external assault, when overexpressed, it can be injurious to the patient, particularly in a cohort of patients in which there is a heightened and uncontrolled response. The inflammatory response develops in these patients to an exaggerated level resulting in an autoimmune injury and may lead to poor postoperative outcomes (systemic inflammatory response syndrome and multi-organs failure). Previous studies by this group have suggested a correlation between the level of IL6 measured in patient’s blood before surgery and after polymeric activation and the observed inflammatory response during surgery. Based upon these findings, the present work is aimed at using this response to develop a test which can be used prior to the open heart surgery to identify the high-risk patients before their operation. The work will be accomplished via three main clinical phases including some pilot in-vitro studies, device development and clinical investigation. Current findings from studies using animal blood, employing DEHP and DEHP plasticized PVC materials as the activator, support the earlier results in patient samples. Having established this relationship, ongoing work will focus on developing an activated lateral flow strip technology as a screening device for heightened inflammatory propensity.

Keywords: cardiopulmonary bypass, cytokines, inflammatory response, overexpression

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989 Influence of Improved Roughage Quality and Period of Meal Termination on Digesta Load in the Digestive Organs of Goats

Authors: Rasheed A. Adebayo, Mehluli M. Moyo, Ignatius V. Nsahlai

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Ruminants are known to relish roughage for productivity but the effect of its quality on digesta load in rumen, omasum, abomasum and other distal organs of the digestive tract is yet unknown. Reticulorumen fill is a strong indicator for long-term control of intake in ruminants. As such, the measurement and prediction of digesta load in these compartments may be crucial to productivity in the ruminant industry. The current study aimed at determining the effect of (a) diet quality on digesta load in digestive organs of goats, and (b) period of meal termination on the reticulorumen fill and digesta load in other distal compartments of the digestive tract of goats. Goats were fed with urea-treated hay (UTH), urea-sprayed hay (USH) and non-treated hay (NTH). At the end of eight weeks of a feeding trial period, upon termination of a meal in the morning, afternoon or evening, all goats were slaughtered in random groups of three per day to measure reticulorumen fill and digesta loads in other distal compartments of the digestive tract. Both diet quality and period affected (P < 0.05) the measure of reticulorumen fill. However, reticulorumen fill in the evening was larger (P < 0.05) than afternoon, while afternoon was similar (P > 0.05) to morning. Also, diet quality affected (P < 0.05) the wet omasal digesta load, wet abomasum, dry abomasum and dry caecum digesta loads but did not affect (P > 0.05) both wet and dry digesta loads in other compartments of the digestive tract. Period of measurement did not affect (P > 0.05) the wet omasal digesta load, and both wet and dry digesta loads in other compartments of the digestive tract except wet abomasum digesta load (P < 0.05) and dry caecum digesta load (P < 0.05). Both wet and dry reticulorumen fill were correlated (P < 0.05) with omasum (r = 0.623) and (r = 0.723), respectively. In conclusion, reticulorumen fill of goats decreased by improving the roughage quality; and the period of meal termination and measurement of the fill is a key factor to the quantity of digesta load.

Keywords: digesta, goats, meal termination, reticulo-rumen fill

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988 Spectrophotometric Determination of L-Dopa in Germinated and Non-Germinated Broad Beans (Vicia faba L.) and Chickpea (Cicer aritinum L.)

Authors: Wissame Gouigah, Amina Medellel, Mahmoud Trachi, Djedjiga Benamara, Salem Benamara

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The purpose of this work is to investigate, by UV/VIS spectrophotometry, the distribution of L-dopa, known as precursor of dopamine which is used in the treatment of Parkinson's disease, in broad beans (Vicia faba) (Vf) and chickpea (Cicer aritinum L.) (CA). In the case of Vf, the different organs were analyzed separately: 1) First, in the fresh state: pod (GF), cotyledons (CF), green shell (EF) and placenta (PF) which is the organ through which the seed is attached to the pod, 2) in the dry state (S): peel of the dry seed (ES) and cotyledons (CS), and 3) in the germinated state: peel (EGe), cotyledons (CGe) and germ (GeVf). Results showed that the content of L-dopa is unevenly distributed between different parts of fresh Vf. But the most important result concerns the predominance of L-dopa in placenta with an L-dopa content (~ 60 mg/g of wet weight, ww) sometimes 7-fold higher (p≤0.05) than those of other considered parts of fresh Vf. In the case of CA, the L-dopa concentration in germinated gains was higher than those found in all analyzed Vf organs, excepted PF.

Keywords: broad bean (Vicia faba L.), chickpea (Cicer aritinum L.), L-dopa, Parkinson disease, placenta

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987 Clinical Presentation and Immune Response to Intramammary Infection of Holstein-Friesian Heifers with Isolates from Two Staphylococcus aureus Lineages

Authors: Dagmara A. Niedziela, Mark P. Murphy, Orla M. Keane, Finola C. Leonard

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Staphylococcus aureus is the most frequent cause of clinical and subclinical bovine mastitis in Ireland. Mastitis caused by S. aureus is often chronic and tends to recur after antibiotic treatment. This may be due to several virulence factors, including attributes that enable the bacterium to internalize into bovine mammary epithelial cells, where it may evade antibiotic treatment, or evade the host immune response. Four bovine-adapted lineages (CC71, CC97, CC151 and ST136) were identified among a collection of Irish S. aureus mastitis isolates. Genotypic variation of mastitis-causing strains may contribute to different presentations of the disease, including differences in milk somatic cell count (SCC), the main method of mastitis detection. The objective of this study was to investigate the influence of bacterial strain and lineage on host immune response, by employing cell culture methods in vitro as well as an in vivo infection model. Twelve bovine adapted S. aureus strains were examined for internalization into bovine mammary epithelial cells (bMEC) and their ability to induce an immune response from bMEC (using qPCR and ELISA). In vitro studies found differences in a variety of virulence traits between the lineages. Strains from lineages CC97 and CC71 internalized more efficiently into bovine mammary epithelial cells (bMEC) than CC151 and ST136. CC97 strains also induced immune genes in bMEC more strongly than strains from the other 3 lineages. One strain each of CC151 and CC97 that differed in their ability to cause an immune response in bMEC were selected on the basis of the above in vitro experiments. Fourteen first-lactation Holstein-Friesian cows were purchased from 2 farms on the basis of low SCC (less than 50 000 cells/ml) and infection free status. Seven cows were infected with 1.73 x 102 c.f.u. of the CC97 strain (Group 1) and another seven with 5.83 x 102 c.f.u. of the CC151 strain (Group 2). The contralateral quarter of each cow was inoculated with PBS (vehicle). Clinical signs of infection (temperature, milk and udder appearance, milk yield) were monitored for 30 days. Blood and milk samples were taken to determine bacterial counts in milk, SCC, white blood cell populations and cytokines. Differences in disease presentation in vivo between groups were observed, with two animals from Group 2 developing clinical mastitis and requiring antibiotic treatment, while one animal from Group 1 did not develop an infection for the duration of the study. Fever (temperature > 39.5⁰C) was observed in 3 animals from Group 2 and in none from Group 1. Significant differences in SCC and bacterial load between groups were observed in the initial stages of infection (week 1). Data is also being collected on cytokines and chemokines secreted during the course of infection. The results of this study suggest that a strain from lineage CC151 may cause more severe clinical mastitis, while a strain from lineage CC97 may cause mild, subclinical mastitis. Diversity between strains of S. aureus may therefore influence the clinical presentation of mastitis, which in turn may influence disease detection and treatment needs.

Keywords: Bovine mastitis, host immune response, host-pathogen interactions, Staphylococcus aureus

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986 Computational Approaches to Study Lineage Plasticity in Human Pancreatic Ductal Adenocarcinoma

Authors: Almudena Espin Perez, Tyler Risom, Carl Pelz, Isabel English, Robert M. Angelo, Rosalie Sears, Andrew J. Gentles

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Pancreatic ductal adenocarcinoma (PDAC) is one of the most deadly malignancies. The role of the tumor microenvironment (TME) is gaining significant attention in cancer research. Despite ongoing efforts, the nature of the interactions between tumors, immune cells, and stromal cells remains poorly understood. The cell-intrinsic properties that govern cell lineage plasticity in PDAC and extrinsic influences of immune populations require technically challenging approaches due to the inherently heterogeneous nature of PDAC. Understanding the cell lineage plasticity of PDAC will improve the development of novel strategies that could be translated to the clinic. Members of the team have demonstrated that the acquisition of ductal to neuroendocrine lineage plasticity in PDAC confers therapeutic resistance and is a biomarker of poor outcomes in patients. Our approach combines computational methods for deconvolving bulk transcriptomic cancer data using CIBERSORTx and high-throughput single-cell imaging using Multiplexed Ion Beam Imaging (MIBI) to study lineage plasticity in PDAC and its relationship to the infiltrating immune system. The CIBERSORTx algorithm uses signature matrices from immune cells and stroma from sorted and single-cell data in order to 1) infer the fractions of different immune cell types and stromal cells in bulked gene expression data and 2) impute a representative transcriptome profile for each cell type. We studied a unique set of 300 genomically well-characterized primary PDAC samples with rich clinical annotation. We deconvolved the PDAC transcriptome profiles using CIBERSORTx, leveraging publicly available single-cell RNA-seq data from normal pancreatic tissue and PDAC to estimate cell type proportions in PDAC, and digitally reconstruct cell-specific transcriptional profiles from our study dataset. We built signature matrices and optimized by simulations and comparison to ground truth data. We identified cell-type-specific transcriptional programs that contribute to cancer cell lineage plasticity, especially in the ductal compartment. We also studied cell differentiation hierarchies using CytoTRACE and predict cell lineage trajectories for acinar and ductal cells that we believe are pinpointing relevant information on PDAC progression. Collaborators (Angelo lab, Stanford University) has led the development of the Multiplexed Ion Beam Imaging (MIBI) platform for spatial proteomics. We will use in the very near future MIBI from tissue microarray of 40 PDAC samples to understand the spatial relationship between cancer cell lineage plasticity and stromal cells focused on infiltrating immune cells, using the relevant markers of PDAC plasticity identified from the RNA-seq analysis.

Keywords: deconvolution, imaging, microenvironment, PDAC

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