Search results for: immune dysfunction
Commenced in January 2007
Frequency: Monthly
Edition: International
Paper Count: 1078

Search results for: immune dysfunction

778 Epstein-Barr Virus-associated Diseases and TCM Syndromes Types: In Search for Correlation

Authors: Xu Yifei, Le Yining, Yang Qingluan, Tu Yanjie

Abstract:

Objective: This study aims to investigate the distribution features of Traditional Chinese Medicine (TCM) syndromes and syndrome elements in Epstein-Barr virus-associated diseases and then explores the relations between TCM syndromes or syndrome elements and laboratory indicators of Epstein-Barr virus-associated diseases. Methods: A cross-sectional study of 70 patients with EBV infection was described. We assessed the diagnostic information and laboratory indicators of these patients from Huashan Hospital Affiliated to Fudan University between November 2017 and July 2019. The disease diagnosis and syndrome differentiation were based on the diagnostic criteria of EBV-associated diseases and the theory of TCM respectively. Confidence correlation analysis, logistic regression analysis, cluster analysis, and the Sankey diagram were used to analyze the correlation between the data. Results: The differentiation of the 4 primary TCM syndromes in the collected patients was correlated with the indexes of immune function, liver function, inflammation, and anemia, especially the relationship between Qifen syndrome and high lactic acid dehydrogenase level. The common 11 TCM syndrome elements were associated with the increased CD3+ T cell rate, low hemoglobin level, high procalcitonin level, high lactic acid dehydrogenase level, and low albumin level. Conclusion: The changes in immune function indexes, procalcitonin, and liver function-related indexes in patients with EBV-associated diseases were consistent with the evolution law of TCM syndromes. This study provides a reference for judging the pathological stages of these kinds of diseases, predicting their prognosis, and guiding subsequent treatment strategies based on TCM syndrome type.

Keywords: EBV-associated diseases, traditional Chinese medicine syndrome, syndrome element, diagnostics

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777 The Effect of Modified Posterior Shoulder Stretching Exercises on Posterior Shoulder Tightness, Shoulder Pain, and Dysfunction in Patients with Subacromial Impingement

Authors: Ozge Tahran, Sevgi Sevi Yesilyaprak

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Objective: The aim of the study was to investigate the effect of the Wilk’s modified two different stretching exercises on posterior shoulder tightness, pain, and dysfunction in patients with subacromial impingement syndrome (SIS). Method: This study was carried out on 67 patients who have more than 15° difference in shoulder internal rotation range of motion between two sides and had been diagnosed as SIS. Before treatment, all patients were randomly assigned into three groups. Standard physiotherapy programme was applied to the Group 3 (n=23), standard physiotherapy program with Wilk’s modified cross-body stretching exercises were applied to Group 1 (n=22), and standard physiotherapy program with Wilk’s modified sleeper stretching exercises were applied to Group 2 (n= 23). All the patients received 20 sessions of physiotherapy during 4 weeks, 5 days in a week by a physiotherapist. The patients continued their exercises at home at the weekends. Pain severity, shoulder rotation range of motion, posterior shoulder tightness, upper extremity functionality with Constant and Murley Score (CMS) and disability level with The Disabilities of the Arm, Shoulder and Hand Score (QuickDASH) were evaluated before and after physiotherapy programme. Results: Before treatment, demographic and anthropometric characteristics were similar in groups and there was no statistical difference (p > 0.05). It was determined that pain severity decreased, shoulder rotation range of motion, posterior shoulder tightness, upper extremity functionality, and disability were improved after physiotherapy in both groups (p < 0.05). Group 1 and 2 had better results in terms of reduction of pain severity during activity, increase in shoulder rotation range of motion, posterior shoulder mobility and upper extremity functionality and improvement in upper extremity disability, compared to Group 3 (p < 0.05). Conclusion: Modified posterior shoulder stretching exercises in addition to standard physiotherapy programme is more effective for reduction of pain during activity, to improve shoulder rotation range of motion, posterior shoulder mobility, and upper extremity functionality in patients with SIS compared to standard physiotherapy programme alone.

Keywords: modified posterior shoulder stretching exercises, posterior shoulder tightness, shoulder complex, subacromial impingement syndrome

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776 Brain Derived Neurotrophic Factor (BDNF) Down Regulation in Peritoneal Carcinomatosis Patients

Authors: Awan A. Zaima, Tanvieer Ayesha, Mirshahi Shahsoltan, Pocard Marc, Mirshahi Massoud

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Brain-derived neurotrophic factor (BDNF) is described as a factor helping to support the survival of existing neurons by involving the growth and differentiation of new neurons and synapses. Cancer diagnosis impacts the mental health, and in consequences, depression arise eventually hinders recovery and disrupts the quality of life and surviving chances of patients. The focus of this study is to hint upon a prospective biomarker as a promising diagnostic tool for an early indicator/predictor of depression prevalence in cancer patients for better care and treatment options. The study aims to analyze peripheral biomarkers from neuro immune axis (BDNF, IL21 as a NK cell activator) using co-relation approach. Samples were obtained from random non cancer candidates and advanced peritoneum carcinomatosis patients with 25% pseudomyxoma, 21% Colon cancer,19% stomach cancer, 10% ovarian cancer, 8% appendices cancer, and 10% other area of peritoneum cancer patients. Both groups of the study were categorized by gender and age, with a range of 18 to 86 years old. Biomarkers were analyzed in collected plasma by performing multiplex sandwich ELISA system. Data were subjected to statistical analysis for the assessment of the correlation. Our results demonstrate that BNDF and IL 21 down regulated significantly in patient groupas compared to non-cancer candidates (ratio of patients/normalis 2.57 for BNDF and 1.32 for IL21). This preliminary investigation suggested that the neuro immune biomarkers are down regulated in carcinomatosis patients and can be associated with cancer expansion and cancer genesis. Further studies on larger cohort are necessary to validate this hypothesis.

Keywords: biomarkers, depression, peritoneum carcinoma, BNDF, IL21

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775 Ribosomal Protein S4 Gene: Exploring the Presence in Syrian Strain of Leishmania Tropica Genome, Sequencing it and Evaluating Immune Response of pCI-S4 DNA Vaccine

Authors: Alyaa Abdlwahab

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Cutaneous leishmaniasis represents a serious health problem in Syria; this problem has become noticeably aggravated after the civil war in the country. Leishmania tropica parasite is the main cause of cutaneous leishmaniasis in Syria. In order to control the disease, we need an effective vaccine against leishmania parasite. DNA vaccination remains one of the favorable approaches that have been used to face cutaneous leishmaniasis. Ribosomal protein S4 is responsible for important roles in Leishmania parasite life. DNA vaccine based on S4 gene has been used against infections by many species of Leishmania parasite but leishmania tropica parasite, so this gene represents a good candidate for DNA vaccine construction. After proving the existence of ribosomal protein S4 gene in a Syrian strain of Leishmania tropica (LCED Syrian 01), sequencing it and cloning it into pCI plasmid, BALB/C mice were inoculated with pCI-S4 DNA vaccine. The immune response was determined by monitoring the lesion progression in inoculated BALB/C mice for six weeks after challenging mice with Leishmania tropica (LCED Syrian 01) parasites. IL-12, IFN-γ, and IL-4 were quantified in draining lymph nodes (DLNa) of the immunized BALB/C mice by using the RT-qPCR technique. The parasite burden was calculated in the final week for the footpad lesion and the DLNs of the mice. This study proved the existence and the expression of the ribosomal protein S4 gene in Leishmania tropica (LCED Syrian 01) promastigotes. The sequence of ribosomal protein cDNA S4 gene was determined and published in Genbank; the gene size was 822 bp. Expression was also demonstrated at the level of cDNA. Also, this study revealed that pCI-S4 DNA vaccine induces TH1\TH2 response in immunized mice; this response prevents partially developing a dermal lesion of Leishmania.

Keywords: ribosomal protein S4, DNA vaccine, Leishmania tropica, BALB\c

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774 Primary Cryptococcal Pneumonia in an HIV Positive Filipino Patient

Authors: Mark Andrew Tu, Raymond Olazo, Cybele Abad

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Cryptococcosis is an invasive infection most commonly found in patients who are immuno compromised. However, patients with this infection usually present with meningitis and rarely pulmonary infection in isolation. We present a case of a Filipino HIV patient who developed cryptococcal pneumonia without meningitis.

Keywords: Cryptococcal Pneumonia, HIV, Filipino, immune system

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773 Effect of Coated Sodium Butyrate (CM3000®) On Zootechnical Performance, Immune Status and Necrotic Enteritis After Experimental Infection of Broiler Chickens

Authors: Mohamed Ahmed Tony, Mohamed Hamoud

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The present study was conducted to determine the effect of commercially coated slow-release sodium butyrate (CM3000®) as a feed additive on zootechnical performance, immune status and Clostridium perfringens severity after experimental infection. Three hundred 1-d-old broiler chicks (Cobb 500) were randomly distributed into 3 treatment groups (4 replicates each) using 25 chicks per replicate on floor pens. Control (C) birds were offered non-supplemented basal diets. Treatments 1 and 2 (T1 and T2) were fed diets containing CM3000® at 300 and 500 g/ton feed, respectively, during the entire experimental period (35 days). Feed and water were offered ad-libitum. Feed consumption and body weight were recorded weekly to calculate body weight gain and feed conversion. Blood samples were collected to evaluate the immune status of the birds against Newcastle disease vaccines using HI test. At the end of the experimental period, 20 birds were chosen randomly from each group (5 birds from each pen) to compare carcass yield. At day 16 of age 20 birds from each group (5 birds/replicate) were bacteriologically examined and proved to be free from Clostridium perfringens. The isolated birds were challenged orally with 1 ml buffer containing 106 CFU/ml Clostridium perfringens local isolate and prepared from necrotic enteritis (NE) diseased farms. Birds were observed on a regular basis daily for any signs of NE. Birds that died in the challenged group were necropsied to determine the cause of death. On day 28 of age, the surviving chickens were killed by cervical dislocation and necropsied immediately. Intestinal tracts were removed and intestinal lesions were scored. Tissue samples of the duodenum, jejunum, ileum and cecum for histopathological examination were collected. All collected data were statistically analyzed using IBM SPSS® version 19 software for personal computers. Means were compared by one-way ANOVA (P<0.05) followed by the Duncan Post Hoc test. The results revealed that body weight gain was significantly (P<0.05) improved in chicks fed on both doses of CM3000® compared to the control one. Final body weight gain in T1 and T2 were 2064.94 and 2141.37 g/bird, respectively, while in the control group, the weight gain showed 1952.78 g/bird. In addition, supplementation of diets with CM3000® increased significantly feed intake (P<0.05). Total feed intake in T1 and T2 were 3186.32 and 3273.29 g/bird, respectively; however, feed intake in the control group recorded 3081.95 g/bird. The best feed conversion was recorded in T2 group (1.53). Feed conversion in the control and T1 groups were 1.58 and 1.54, respectively. Dressing percentage, liver weights and the other carcasses yields were not different between treatments. The butyrate significantly enhanced immune responses measured against Newcastle disease vaccines. Sodium butyrate significantly reduced NE lesions and healthy improved the intestinal tissues in the samples collected from T1 and T2-challenged chickens versus those collected from the control group. In conclusion, exogenous administration of slow-release butyrate (CM3000®) is capable of improving performance, enhancing immunity and NE disease resistance in broiler chickens.

Keywords: sodium butyrate, broiler chicken, zootechnical performance, immunity, necrotic enteritis

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772 Maresin Like 1 Treatment: Curbing the Pathogenesis of Behavioral Dysfunction and Neurodegeneration in Alzheimer's Disease Mouse Model

Authors: Yan Lu, Song Hong, Janakiraman Udaiyappan, Aarti Nagayach, Quoc-Viet A. Duong, Masao Morita, Shun Saito, Yuichi Kobayashi, Yuhai, Zhao, Hongying Peng, Nicholas B. Pham, Walter J Lukiw, Christopher A. Vuong, Nicolas G. Bazan

Abstract:

Aims: Neurodegeneration and behavior dysfunction occurs in patients with Alzheimer's Disease (AD), and as the disease progresses many patients develop cognitive impairment. 5XFAD mouse model of AD is widely used to study AD pathogenesis and treatment. This study aimed to investigate the effect of maresin like 1 (MaR-L1) treatment in AD pathology using 5XFAD mice. Methods: We tested 12-month-old male 5XFAD mice and wild type control mice treated with MaR-L1 in a battery of behavioral tasks. We performed open field test, beam walking test, clasping test, inverted grid test, acetone test, marble burring test, elevated plus maze test, cross maze test and novel object recognition test. We also studied neuronal loss, amyloid β burden, and inflammation in the brains of 5XFAD mice using immunohistology and Western blotting. Results: MaR-L1 treatment to the 5XFAD mice showed improved cognitive function of 5XFAD mice. MaR-L1 showed decreased anxiety behavior in open field test and marble burring test, increased muscular strength in the beam walking test, clasping test and inverted grid test. Cognitive function was improved in MaR-L1 treated 5XFAD mice in the novel object recognition test. MaR-L1 prevented neuronal loss and aberrant inflammation. Conclusion: Our finding suggests that behavioral abnormalities were normalized by the administration of MaR-L1 and the neuroprotective role of MaR-L1 in the AD. It also indicates that MaR-L1 treatment is able to prevent and or ameliorate neuronal loss and aberrant inflammation. Further experiments to validate the results are warranted using other AD models in the future.

Keywords: Alzheimer's disease, motor and cognitive behavior, 5XFAD mice, Maresin Like 1, microglial cell, astrocyte, neurodegeneration, inflammation, resolution of inflammation

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771 Probable Future Weapon to Turn down Malnutrition in Women and Adolescent Girls

Authors: Manali Chakraborty

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In the developing countries the most prevalent pathological state is malnutrition and under nutrition due to deficiency of essential nutrients. This condition is more seen between the woman population, especially in the adolescent girls. It is causing childhood deaths along with others cognitive, degenerative diseases. Born of low weight babies and stillbirth are also major problems associated with the malnutrition. Along with the increased level of population, people not only should concern about their quantity of food but also for the quality of the food to be healthy. Lethargy, depression caused due to iron deficiency often quoted as normal or unimportant issues. Children of malnourished women are more likely associated with cognitive impairment, immune dysfunction leading to a higher risk of being attacked by diseases. Malnourishment also affects the productivity of women. Low social status, lack of proper nutritional education is an important cause of this nutrient deficiency among women. Iron deficiency and anemia are mostly famous nutritional deficiencies among women worldwide. Mostly women from below poverty lined area are anemic due to less consumption of iron-rich foods or having foods that might inhibit the iron absorption. Growing females like adolescents or lactating females need more iron supply. Less supplement causes iron deficiency. Though malaria also might cause anemia, it is more likely endemic one in some specific areas. Folate deficiency in females also may cause neurological defects in the infants. Other Vitamin B, A deficiencies along with low iodine level is also noted in malnourished women. According to tradition, still in some areas in developing countries females have their food at the end. According to some survey and collected data, these females are often into the risk zone of being malnourished. Regularly they have the very lesser amount, or food sometimes may start to lose its nutrients. Women are the one who maintains the responsibility to cook foods in the home. Lack of proper nutritional education and proper food preparation not only make those foods lose the nutrients leading to impairment in proper nutrient intake of the family members but also impairing own nutritional status. Formulation and development of food products from iron or other nutrient affluent sources viz., traditional herbs can be helpful in the prevention of such malnutrition condition in females. Keeping low cost and smooth maintenance of the food product development from the natural sources like pulses, cereals or other vegetation also can be beneficial to sustain socio-economic condition. Consumption of such kind of foodstuff is much healthier rather than taking continuous supplements like capsules. Utilization of proper scientific and cost-effective techniques for this food product development and their distribution among rural women population might be an enormous initiative.

Keywords: anemia, food supplements, malnutrition, rural places, women population

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770 Antifungal Susceptibility of Saprolegnia parasitica Isolated from Rainbow Trout and Its Host Pathogen Interaction in Zebrafish Disease Model

Authors: Sangyeop Shin, D. C. M. Kulatunga, S. H. S. Dananjaya, Chamilani Nikapitiya, Jehee Lee, Mahanama De Zoysa

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Saprolegniasis is one of the most devastating fungal diseases in freshwater fish which is caused by species in the genus Saprolegnia including Saprolegnia parasitica. In this study, we isolated the strain of S. parasitica from diseased rainbow trout in Korea. Morphological and molecular based identification confirmed that isolated fungi belong to the member of S. parasitica, supported by its typical fungal features including cotton-like whitish mycelium, zoospores (primary and secondary) and phylogenetic analysis with internal transcribed spacer (ITS) region. Pathogenicity of isolated S. parasitica was developed in embryo, larvae, juvenile and adult zebrafish as a disease model. Up regulation of host genes encoding ZfTnf-α, Zfc-Rel, ZfIl-12, ZfLyz-c, Zfβ-def, and ZfHsp-70 was identified in zebrafish larvae after experimental challenge of S. parasitica showing the host immune responses against the S. parasitica. Survival of the juveniles upon fungal infection might be due to the increased immune protection in the host. Investigation of antifungal susceptibility of S. parasitica with natural lawsone (2-hydroxy-1,4-naphthoquinone) revealed the minimum inhibitory concentration (MIC) and percentage inhibition of radial growth (PIRG %) as 200 µg/mL and 31.8%, respectively. Lawsone was able to change the membrane permeability, and cause irreversible damage and disintegration to the cellular membranes of S. parasitica which might have effect on fungi growth inhibition. Moreover, the mycelium exposed to lawsone (MIC level) changed the transcriptional responses of S. parasitica genes. Overall results indicate that lawsone could be a potential and novel anti-S. parasitica agent for controlling S. parasitica infection.

Keywords: host-pathogen interactions, lawsone, rainbow trout, Saprolegnia parasitica, Saprolegniasis, zebrafish

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769 Prevalence of Diabetes Mellitus Among Human Immune Deficiency Virus-Positive Patients Under Anti-retroviral Attending in Rwanda, a Case Study of University Teaching Hospital of Butare

Authors: Venuste Kayinamura, V. Iyamuremye, A. Ngirabakunzi

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Anti-retroviral therapy (ART) for HIV patient can cause a deficiency in glucose metabolism by promoting insulin resistance, glucose intolerance, and diabetes, diabetes mellitus keep increasing among HIV-infected patients worldwide but there is limited data on levels of blood glucose and its relationship with antiretroviral drugs (ARVs) and HIV-infection worldwide, particularly in Rwanda. A convenient sampling strategy was used in this study and it involved 323 HIV patients (n=323). Patients who are HIV positive under ARVs were involved in this study. The patient’s blood glucose was analyzed using an automated machine or glucometer (COBAS C 311). Data were analyzed using Microsoft Excel and SPSS V. 20.0 and presented in percentages. The highest diabetes mellitus prevalence was 93.33 % in people aged >40 years while the lowest diabetes mellitus prevalence was 6.67% in people aged between 21-and 40 years. The P-value was (0.021). Thus, there is a significant association between age and diabetes occurrence. The highest diabetes mellitus prevalence was 28.2% in patients under ART treatment for more than 10 years, 16.7% were <5years while 20% of patients were on ART treatment between 5-10 years. The P-value here is (0.03), thus the incidence of diabetes is associated with long-term ART use in HIV-infected patients. This study assessed the prevalence of diabetes among HIV-infected patients under ARVs attending the University Teaching Hospital of Butare (CHUB), it shows that the prevalence of diabetes is high in HIV-infected patients under ARTs. This study found no significant relationship between gender and diabetes mellitus growth. Therefore, regular assessment of diabetes mellitus especially among HIV-infected patients under ARVs is highly recommended to control other health issues caused by diabetes mellitus.

Keywords: anti-retroviral, diabetes mellitus, antiretroviral therapy, human immune deficiency virus

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768 Study of a Cross-Flow Membrane to a Kidney Encapsulation Engineering Structures for Immunosuppression Filter

Authors: Sihyun Chae, Ryoto Arai, Waldo Concepcion, Paula Popescu

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The kidneys perform an important role in the human hormones that regulate the blood pressure, produce an active form of vitamin D and control the production of red blood cells. Kidney disease can cause health problems, such as heart disease. Also, increase the chance of having a stroke or heart attack. There are mainly to types of treatments for kidney disease, dialysis, and kidney transplant. For a better quality of life, the kidney transplant is desirable. However, kidney transplant can cause antibody reaction and patients’ body would be attacked by immune system of their own. For solving that issue, patients with transplanted kidney always take immunosuppressive drugs which can hurt kidney as side effects. Patients willing to do a kidney transplant have a waiting time of 3.6 years in average searching to find an appropriate kidney, considering there are almost 96,380 patients waiting for kidney transplant. There is a promising method to solve these issues: bioartificial kidney. Our membrane is specially designed with unique perforations capable to filter the blood cells separating the white blood cells from red blood cells. White blood cells will not pass through the encapsulated kidney preventing the immune system to attack the new organ and eliminating the need of a matching donor. It is possible to construct life-time long encapsulation without needing pumps or a power supply on the cell’s separation method preventing futures surgeries due the Cross-Channel Flow inside the device. This technology allows the possibility to use an animal kidney, prevent cancer cells to spread through the body, arm and leg transplants in the future. This project aims to improve the quality of life of patients with kidney disease.

Keywords: kidney encapsulation, immunosuppression filter, leukocyte filter, leukocyte

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767 Impact of Helicobacter pylori Infection on Colorectal Adenoma-Colorectal Carcinoma Sequence

Authors: Jannis Kountouras, Nikolaos Kapetanakis, Stergios A. Polyzos, Apostolis Papaeftymiou, Panagiotis Katsinelos, Ioannis Venizelos, Christina Nikolaidou, Christos Zavos, Iordanis Romiopoulos, Elena Tsiaousi, Evangelos Kazakos, Michael Doulberis

Abstract:

Background & Aims: Helicobacter pylori infection (Hp-I) has been recognized as a substantial risk agent involved in gastrointestinal (GI) tract oncogenesis by stimulating cancer stem cells (CSCs), oncogenes, immune surveillance processes, and triggering GI microbiota dysbiosis. We aimed to investigate the possible involvement of active Hp-I in the sequence: chronic inflammation–adenoma–colorectal cancer (CRC) development. Methods: Four pillars were investigated: (i) endoscopic and conventional histological examinations of patients with CRC, colorectal adenomas (CRA) versus controls to detect the presence of active Hp-I; (ii) immunohistochemical determination of the presence of Hp; expression of CD44, an indicator of CSCs and/or bone marrow-derived stem cells (BMDSCs); expressions of oncogene Ki67 and anti-apoptotic Bcl-2 protein; (iii) expression of CD45, indicator of immune surveillance locally (assessing mainly T and B lymphocytes locally); and (iv) correlation of the studied parameters with the presence or absence of Hp-I. Results: Among 50 patients with CRC, 25 with CRA, and 10 controls, a significantly higher presence of Hp-I in the CRA (68%) and CRC group (84%) were found compared with controls (30%). The presence of Hp-I with accompanying immunohistochemical expression of CD44 in biopsy specimens was revealed in a high proportion of patients with CRA associated with moderate/severe dysplasia (88%) and CRC patients with moderate/severe degree of malignancy (91%). Comparable results were also obtained for Ki67, Bcl-2, and CD45 immunohistochemical expressions. Concluding Remarks: Hp-I seems to be involved in the sequence: CRA – dysplasia – CRC, similarly to the upper GI tract oncogenesis, by several pathways such as the following: Beyond Hp-I associated insulin resistance, the major underlying mechanism responsible for the metabolic syndrome (MetS) that increase the risk of colorectal neoplasms, as implied by other Hp-I related MetS pathologies, such as non-alcoholic fatty liver disease and upper GI cancer, the disturbance of the normal GI microbiota (i.e., dysbiosis) and the formation of an irritative biofilm could contribute to a perpetual inflammatory upper GIT and colon mucosal damage, stimulating CSCs or recruiting BMDSCs and affecting oncogenes and immune surveillance processes. Further large-scale relative studies with a pathophysiological perspective are necessary to demonstrate in-depth this relationship.

Keywords: Helicobacter pylori, colorectal cancer, colorectal adenomas, gastrointestinal oncogenesis

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766 Effect of Dietary Graded Levels of L-Theanine on Growth Performance, Carcass Traits, Meat Quality, and Immune Response of Broilers

Authors: Muhammad Saeed, Sun Chao

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L-theanine is water soluble non-proteinous amino acid found in green tea leaves. Despite the availability of abundant literature on green tea, studies on the use of L-theanine as an additive in animals especially broilers are scanty. The objective of this study was to evaluate the effectiveness of different dietary levels of L-theanine on growth performance, meat quality, growth, immune response and blood chemistry in broilers. A total of 400 day-old chicks were randomly divided into four treatment groups (A, B, C, and D) using a complete randomized design. Treatments were as follows: A; control (basal diet), B; basal diet+100 mg L-theanine / kg diet, C; basal diet+ 200 mg L-theanine / kg diet, and D; basal diet+ 300 mg L-theanine / kg diet. Results revealed that intermediate level of L-theanine (200 mg/ kg diet, group C) showed better results in terms of BWG, FC, and FCR compared with control and other L-theanine levels. The live weight eviscerated weight and gizzard weight was higher in all L-theanine levels as compared to that of the control group. The heaviest (P > 0.05) spleen and bursa were found in group C (200 mg L-theanine / kg diet). Analysis of meat colors according to yellowness (b*), redness (a*), and lightness (L*) showed significantly higher values of a* and b* in L-theanine groups. Supplementing broiler diet with L-theanine minimized (P=0.02) total cholesterol contents in serum. Further analysis revealed , lower mRNA expression of TNF-α and IL-6 in thymus and IFN- γ and IL-2 in spleen was observed in L-theanine group It is concluded that supplementation of L-theanine at 200mg/kg diet showed better results in terms of performance and it could be utilized as a natural feed additive alternative to antibiotics to improve overall performance of broilers. Increasing the levels up to 300 mg L-theanine /kg diet may has deleterious effects on performance and other health aspects.

Keywords: blood chemistry, broilers growth, L-theanine, meat quality

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765 Influence of Cyperus Rotundus Active Principles Inhibit Viral Multiplication and Stimulate Immune System in Indian White Shrimp Fenneropenaeus Indicus against White Spot Syndrome Virus Infection

Authors: Thavasimuthu Citarasu, Mariavincent Michaelbabu, Vikram Vakharia

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The rhizome of Java grass, Cyperus rotundus was extracted different organic polar and non-polar solvents and performed the in vitro antiviral and immunostimulant activities against White Spot Syndrome Virus (WSSV) and Vibrio harveyi respectively. Based on the initial screening the ethyl acetate extract of C. rotundus was strong activities and further it was purified through silica column chromatography and the fractions were screened again for antiviral and immunostimulant activity. Among the different fractions screened against the WSSV and V. harveyi, the fractions, F-III to FV had strong activities. In order to study the in vivo influence of C. rotundus, the fractions (F-III to FV) were pooled and delivered to the F. indicus through artificial feed for 30 days. After the feeding trail the experimental and control diet fed F. indicus were challenged with virulent WSSV and studied the survival, molecular diagnosis, biochemical, haematological and immunological parameters. Surprisingly, the pooled fractions (F-III to FV) incorporated diets helped to significantly (P < 0.01) suppressed viral multiplication, showed significant (P < 0.01) differences in protein and glucose levels, improved total haemocyte count (THC), coagulase activity, significantly increased (P < =0.001) prophenol oxidase and intracellular superoxide anion production compared to the control shrimps. Based on the results, C. rotundus extracts effectively suppressed WSSV multiplication and improve the immune system in F. indicus against WSSV infection and this knowledge will helps to develop novel drugs from C. rotundus against WSSV.

Keywords: antiviral drugs, cyperus rotundus, fenneropenaeus indicus, WSSV

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764 Clinical Evaluation of Neutrophil to Lymphocytes Ratio and Platelets to Lymphocytes Ratio in Immune Thrombocytopenic Purpura

Authors: Aisha Arshad, Samina Naz Mukry, Tahir Shamsi

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Background: Immune thrombocytopenia (ITP) is an autoimmune disorder. Besides platelets counts, immature platelets fraction (IPF) can be used as tool to predict megakaryocytic activity in ITP patients. The clinical biomarkers like Neutrophils to lymphocytes ratio (NLR) and platelet to lymphocytes ratio(PLR) predicts inflammation and can be used as prognostic markers.The present study was planned to assess the ratios in ITP and their utility in predicting prognosis after treatment. Methods: A total of 111 patients of ITP with same number of healthy individuals were included in this case control study during the period of January 2015 to December 2017.All the ITP patients were grouped according to guidelines of International working group of ITP. A 3cc blood was collected in EDTA tube and blood parameters were evaluated using Sysmex 1000 analyzer.The ratios were calculated by using absolute counts of Neutrophils,Lymphocytes and platelets.The significant (p=<0.05) difference between ITP patients and healthy control groups was determined by Kruskal wallis test, Dunn’s test and spearman’s correlation test was done using SPSS version 23. Results: The significantly raised total leucocytes counts (TLC) and IPF along with low platelets counts were observed in ITP patients as compared to healthy controls.In ITP groups,very low platelet count with median and IQR of 2(3.8)3x109/l with highest mean and IQR IPF 25.4(19.8)% was observed in newly diagnosed ITP group. The NLR was high with prognosis of disease as higher levels were observed in P-ITP. The PLR was significantly low in ND-ITP ,P-ITP, C-ITP, R-ITP and compared to controls with p=<0.001 as platelet were less in number in all ITP patients. Conclusion: The IPF can be used in evaluation of bone marrow response in ITP. The simple, reliable and calculated NLR and PLR ratios can be used in predicting prognosis and response to treatment in ITP and to some extend the severity of disease.

Keywords: neutrophils, platelets, lymphocytes, infection

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763 Study of Mixing Conditions for Different Endothelial Dysfunction in Arteriosclerosis

Authors: Sara Segura, Diego Nuñez, Miryam Villamil

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In this work, we studied the microscale interaction of foreign substances with blood inside an artificial transparent artery system that represents medium and small muscular arteries. This artery system had channels ranging from 75 μm to 930 μm and was fabricated using glass and transparent polymer blends like Phenylbis(2,4,6-trimethylbenzoyl) phosphine oxide, Poly(ethylene glycol) and PDMS in order to be monitored in real time. The setup was performed using a computer controlled precision micropump and a high resolution optical microscope capable of tracking fluids at fast capture. Observation and analysis were performed using a real time software that reconstructs the fluid dynamics determining the flux velocity, injection dependency, turbulence and rheology. All experiments were carried out with fully computer controlled equipment. Interactions between substances like water, serum (0.9% sodium chloride and electrolyte with a ratio of 4 ppm) and blood cells were studied at microscale as high as 400nm of resolution and the analysis was performed using a frame-by-frame observation and HD-video capture. These observations lead us to understand the fluid and mixing behavior of the interest substance in the blood stream and to shed a light on the use of implantable devices for drug delivery at arteries with different Endothelial dysfunction. Several substances were tested using the artificial artery system. Initially, Milli-Q water was used as a control substance for the study of the basic fluid dynamics of the artificial artery system. However, serum and other low viscous substances were pumped into the system with the presence of other liquids to study the mixing profiles and behaviors. Finally, mammal blood was used for the final test while serum was injected. Different flow conditions, pumping rates, and time rates were evaluated for the determination of the optimal mixing conditions. Our results suggested the use of a very fine controlled microinjection for better mixing profiles with and approximately rate of 135.000 μm3/s for the administration of drugs inside arteries.

Keywords: artificial artery, drug delivery, microfluidics dynamics, arteriosclerosis

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762 Morphological Variation of the Mesenteric Lymph Node in Dromedary Camels: The Impact of Rearing Systems

Authors: Khenenou Tarek, Mohamed Amine Fares, Djallal Eddine Rahmoun

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The study intends to evaluate the morphological changes in the mesenteric lymph nodes of dromedaries in different rearing systems. we aimed to evaluate the adaptative behavior of the animal’s immune system with environmental variations, and to conduct a comparative analysis on the morphological features of the mesenteric lymph node of the one-humped camel (Camelus dromedarius) in the region of El Oued, with two different rearing systems, with different practices and different purposes. The study was conducted using histo-morphometric techniques to analyze the morphological features of the mesenteric lymph node of the one-humped camel (Camelus dromedarius) in the region of El Oued. Two groups of dromedaries were used in the study, one group raised in a free-roaming housing system and another group raised in a restricted-roaming housing system. The results revealed that there were significant differences between the two groups in terms of active follicle ratio and size and also the cellular population of functional zones. Animals living and roaming outside the farm barriers were more exposed to pathogens, which leads to the installation of an adaptative process, whereas the animals living under restricted-roaming housing system were not exposed to pathogens. This study indicated that the adaptative behavior of the animal’s immune system with environmental variations is the functional translation of morphological changes. The obtained findings revealed that the morphological features of the mesenteric lymph node of the one-humped camel (Camelus dromedarius) in the region of El Oued are directly linked to the rearing system practices

Keywords: adaptative behavior, dromedary, lymph node, morphology, rearing systems

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761 Manganese Contamination Exacerbates Reproductive Stress in a Suicidally-Breeding Marsupial

Authors: Ami Fadhillah Amir Abdul Nasir, Amanda C. Niehaus, Skye F. Cameron, Frank A. Von Hippel, John Postlethwait​, Robbie S. Wilson

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For suicidal breeders, the physiological stresses and energetic costs of breeding are fatal. Environmental stressors such as pollution should compound these costs, yet suicidal breeding is so rare among mammals that this is unknown. Here, we explored the consequences of metal contamination to the health, aging and performance of endangered, suicidally-breeding northern quolls (Dasyurus hallucatus) living near an active manganese mine on Groote Eylandt, Northern Territory, Australia. We found respirable manganese dust at levels exceeding international recommendations even 20km from mining sites and substantial accumulation of manganese within quolls’ hair, testes, and in two brain regions—the neocortex and cerebellum, responsible for sensory perception and motor function, respectively. Though quolls did not differ in sprint speeds, motor skill, or manoeuvrability, those with higher accumulation of manganese crashed at lower speeds during manoeuvrability tests, indicating a potential effect on sight or cognition. Immune function and telomere length declined over the breeding season, as expected with ageing, but manganese contamination exacerbated immune declines and suppressed cortisol. Unexpectedly, male quolls with higher levels of manganese had longer telomeres, supporting evidence of unusual telomere dynamics among Dasyurids—though whether this affects their lifespan is unknown. We posit that sublethal contamination via pollution, mining, or urbanisation imposes physiological costs on wildlife that may diminish reproductive success or survival.

Keywords: ecotoxicology, heavy metal, manganese, telomere length, cortisol, locomotor

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760 Anti-TNF: Possibilities of Rising Anti-Phosphorylcholine Antibodies

Authors: Md. Mizanur Rahman, Anquan Liu, Anna Frostegård, Johan Frostegård

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The role of the human immune system is essential in cardiovascular diseases and atherosclerosis. Activated cells in atherosclerosis produce abundant amounts of cytokines, but the exact mechanisms involved in the effects of these inflammatory cytokines are not clear in atherosclerosis. In a large clinical cohort, we have previously determined that antibodies against phosphorylcholine (anti-PC) are negatively and independently associated with both development of atherosclerosis and also a low risk of cardiovascular disease. Further, we reported that rheumatoid arthritis patients who were non-responders to TNF-inhibitors, where those with low anti-PC levels. Upon anti-TNF treatment, anti-PC levels increased. We, therefore, hypothesised that proinflammatory cytokines such as TNF could play a role in anti-PC regulation. Peripheral blood mononuclear cells (PBMC) were cultured with or without TNF and anti-TNF. The cell supernatants were collected after six days for ELISA measurements. In separate experiments, cells were cultured for 24 hours in both polystyrene plates and ELISPOT plates under a similar condition for ELISA and ELISPOT assays respectively. Total RNA was extracted after 6 hours of cell culture to perform RT-qPCR. Cell viability was confirmed by trypan blue staining and MTT assays. ELISA measurements detected less than 40% of anti-PC in TNF-treated cells, in comparison to control cells, whereas anti-PC production was recovered by anti-TNF treatment. ELISPOT assays showed that TNF suppresses anti-PC production by inhibiting anti-PC producing B-cells. In addition, RT-qPCR and ELISA showed that TNF also has effects also on B-cell activation as BAFF expression was inhibited by TNF treatment. Atherosclerosis is a major cause of cardiovascular diseases, but anti-PC is a protection marker for atherosclerosis development. Our findings show that TNF is a negative regulator of anti-PC production. Immune modulation and rising of anti-PC could be of major significance for the patients.

Keywords: anti-PC, Anti-TNF, atherosclerosis, cardiovascular diseases, phosphorylecholine

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759 The Role of the STAT3 Signaling for Melatonergic Synthetic Pathway in the Rat Pineal Gland

Authors: Simona Moravcova, Jiri Novotny, Zdenka Bendova

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The pineal gland of the vertebrate brain is a circumventricular organ which serves as a major neuroendocrine gland with the primary function of rhythmic secretion of neurohormone melatonin under the control of the hypothalamic suprachiasmatic nucleus (SCN). Soon after the onset of the darkness, the activity of the key rate-limiting enzyme for melatonin synthesis, arylalkylamine N-acetyltransferase (AANAT), raises due to the increased release of norepinephrine from sympathetic neurons terminating on the parenchymal cells where it binds to β-adrenergic receptors. Melatonin codes the length of the night, and it is well recognized for its anti-inflammatory effects. However, to our knowledge, less is known about the effect of the immune system on the melatonin biosynthesis and the precise role of the STAT3 in the signaling pathway leading to the expression of AANAT. Lipopolysaccharide (LPS) is the essential component in the outer surface membrane of gram-negative bacteria and acts as a strong stimulator of natural and innate immunity. STAT3 acts as an important factor in immune response. Here we investigated the effect of LPS on the components of the melatonergic synthetic pathway in the pineal gland. The experiments were performed both in vivo and in vitro. The changes in AANAT activity were determined by radioenzymatic assay. PCR analyses were carried out to detect aa-nat, icer, spi-3 and stat3 gene expression. From our results, it is apparent that the high basal level of phosphorylated forms of STAT3 can be elevated after systemic as well as in vitro administration of LPS. Our experiments have shown that LPS reduces melatonin synthesis, nevertheless, the activity of AANAT was increased. Moreover, the basal level of phosphorylated STAT3 counteracts β-adrenergic receptor-mediated aa-nat gene expression and sustains its own and spi-3 gene expression. In conclusion, LPS can affect immunomodulators such as melatonin in the pineal gland.

Keywords: AANAT, lipopolysaccharide, pineal gland, rat, STAT3

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758 Suggested Role for Neutrophil Extracellular Traps Formation in Ewing Sarcoma Immune Microenvironment

Authors: Rachel Shukrun, Szilvia Baron, Victoria Fidel, Anna Shusterman, Osnat Sher, Netanya Kollender, Dror Levin, Yair Peled, Yair Gortzak, Yoav Ben-Shahar, Revital Caspi, Sagi Gordon, Michal Manisterski, Ronit Elhasid

Abstract:

Ewing sarcoma (EWS) is a highly aggressive cancer with a survival rate of 70–80% for patients with localized disease and under 30% for those with metastatic disease. Tumor-infiltrating neutrophils (TIN) can generate extracellular net-like DNA structures known as neutrophil extracellular traps (NETs). However, little is known about the presence and prognostic significance of tumor-infiltrating NETs in EWS. Herein, we investigated 46 patients diagnosed with EWS and treated in the Tel Aviv Medical Center between 2010 and 2021. TINs and NETs were identified in diagnostic biopsies of EWS by immunofluorescent. In addition, NETs were investigated in neutrophils isolated from peripheral blood samples of EWS patients at diagnosis and following neoadjuvant chemotherapy. The relationships between the presence of TINs and NETs, pathological and clinical features, and outcomes were analyzed. Our results demonstrate that TIN and NETs at diagnosis were higher in EWS patients with metastatic disease compared to those with local disease. High NETs formation at diagnosis predicted poor response to neo-adjuvant chemotherapy, relapse, and death from disease (P < .05). NETs formation in peripheral blood samples at diagnosis was significantly elevated among patients with EWS compared to pediatric controls and decreased significantly following neoadjuvant chemotherapy. In conclusion, NETs formation seems to have a role in the EWS immune microenvironment. Their presence can refine risk stratification, predict chemotherapy resistance and survival, and serve as a therapeutic target in patients with EWS.

Keywords: Ewing sarcoma, tumor microenvironment, neutrophil, neutrophil extracellular traps (NETs), prognosis

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757 Effect of Garlic Extract on Growth Performance and Immune System of Broiler

Authors: Merry Muspita Dyah Utami

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The positive effect of garlic extract have been reported by many studies. It has antibiotical potential, antibacterial, antiviral, antiparasitic, antifungal, and growth promoting. Supplementary garlic for broilers could mediate in getting the bioactive compounds in garlic. The avian bursa must be essential for antibody-mediated immunity. The size of bursa of fabricius must be some sort of endocrine or lymphoid gland associated with growth and sexual development. The research was conducted to evaluate the effects of garlic extract on growth performance and immune system of broiler. Seventy-two day old chick were equally divided into four group, three replication and six chicks each. Group I was control without garlic extract, then garlic extraxt was administrated to the experimental group II, III and IV (2, 4, 6% in ration). The experiment was conducted for three weeks period from day old chick to 21 days. Body weight of broiler were determined at day 1 and 21, feed intake was determined at the same period, feed conversion ratio was calculated accordingly. At 21 day age, four birds per replicate were slaughtered , bursa was collected, weight and calculated as a percentage of live body weight. Mortality was recorded as it occurred and was used to ajust the total number of broiler to determine the total feed intake and feed conversion rasio. Data were expressed as the mean was compare by one way analysis of variance (Anova) follow by Duncan Test, which used to identify differences between groups. A value of P<0.05 was accepted as significance. The body weight, feed conversion rasio, and the weight of bursa of fabricius showed a significant differences, but feed consumption and the percentage of bursa of live body weight were not significantly different (P > 0.05) influenced by dietary treatments. The results of this research, garlic extract has a potential role as natural growth promoter and immunomodulatory system in broiler.

Keywords: garlic extract, growth, immunity, broiler

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756 NLRP3-Inflammassome Participates in the Inflammatory Response Induced by Paracoccidioides brasiliensis

Authors: Eduardo Kanagushiku Pereira, Frank Gregory Cavalcante da Silva, Barbara Soares Gonçalves, Ana Lúcia Bergamasco Galastri, Ronei Luciano Mamoni

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The inflammatory response initiates after the recognition of pathogens by receptors expressed by innate immune cells. Among these receptors, the NLRP3 was associated with the recognition of pathogenic fungi in experimental models. NLRP3 operates forming a multiproteic complex called inflammasome, which actives caspase-1, responsible for the production of the inflammatory cytokines IL-1beta and IL-18. In this study, we aimed to investigate the involvement of NLRP3 in the inflammatory response elicited in macrophages against Paracoccidioides brasiliensis (Pb), the etiologic agent of PCM. Macrophages were differentiated from THP-1 cells by treatment with phorbol-myristate-acetate. Following differentiation, macrophages were stimulated by Pb yeast cells for 24 hours, after previous treatment with specific NLRP3 (3,4-methylenedioxy-beta-nitrostyrene) and/or caspase-1 (VX-765) inhibitors, or specific inhibitors of pathways involved in NLRP3 activation such as: Reactive Oxigen Species (ROS) production (N-Acetyl-L-cysteine), K+ efflux (Glibenclamide) or phagossome acidification (Bafilomycin). Quantification of IL-1beta and IL-18 in supernatants was performed by ELISA. Our results showed that the production of IL-1beta and IL-18 by THP-1-derived-macrophages stimulated with Pb yeast cells was dependent on NLRP3 and caspase-1 activation, once the presence of their specific inhibitors diminished the production of these cytokines. Furthermore, we found that the major pathways involved in NLRP3 activation, after Pb recognition, were dependent on ROS production and K+ efflux. In conclusion, our results showed that NLRP3 participates in the recognition of Pb yeast cells by macrophages, leading to the activation of the NLRP3-inflammasome and production of IL-1beta and IL-18. Together, these cytokines can induce an inflammatory response against P. brasiliensis, essential for the establishment of the initial inflammatory response and for the development of the subsequent acquired immune response.

Keywords: inflammation, IL-1beta, IL-18, NLRP3, Paracoccidioidomycosis

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755 Effects of IMUNO-2865® as Immune Supplement for the Aquaculture Industry

Authors: Ivan Zupan, Tomislav Saric, Suzana Tkalcic

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IMUNO-2865® is a commercially available, β–glucan based, natural hemicellulose compound with proven immunostimulative properties in people, domestic and some aquatic animals. During the experimental feeding trial with IMUNO-2865® in juvenile wild-caught chub under laboratory conditions, supplementation resulted in overall higher growth performance for all experimental groups regardless of the concentration of the added compound. The maximum, 5% concentration of the supplement, resulted in highest weight gain and calculated specific growth rate. In sea bream, as economically most important species in the Mediterranean aquaculture, significant increases in numbers of monocytes and heterophils were observed in the group supplemented with 2.5 % of IMUNO-2865® in the feed. An overall increase of erythrocytes was noted by the end of the experiment, although with variable distribution among groups. Blood Ca++ levels, total proteins, and total NH₃ were significantly higher after 60 days of feeding in all treatment groups compared to the control and remained elevated in the treated group following the secession of supplementation. Superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and serum paraoxonase PON1 (U/L) showed similar trends. All these parameters are playing a significant role in either oxygen supplementation of tissues, or anabolic and catabolic processes that on molecular levels contribute to the overall health and immune-building capacity of cells and tissues. The complete lack of mortality in sea bream and presented increases in cellular, biochemical and oxidative stress parameters in the blood suggest that the IMUNO-2865® represents a safe dietary supplement for in aquaculture, with an overall positive and potentially immunostimulative effect on farmed fish.

Keywords: IMUNO-2865®, β–glucans, Mediterranean aquaculture, fish imunnostimulans

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754 The Morphological Changes of POV in Diabetic Patients and Its Correlation with Changes in Corneal Epithelium, Corneal Nerve, and the Fundus in Using Vivo Confocal Microscopy

Authors: Ji Jiazheng, Wang Jingrao, Jin Xin, Zhang Hong

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Diabetes mellitus is a metabolic disease characterized by high blood sugar. A long-standing hyperglycemic state can lead to various tissue damage. Diabetic retinopathy is the most common and widely studied ocular complication and has become the leading cause of blindness in my country. At the same time, diabetes has profound clinically relevant effects on the cornea, leading to keratopathy and vision-threatening. The cornea is an avascular tissue and is sensitive to hyperglycemia, Keratopathy caused by diabetes is usually chronic, they are called diabetic keratopathy or diabetic neurotrophic keratopathy, leading to several diabetic corneal complications including delayed epithelial wound healing, recurrent erosions, neuropathy, loss of sensitivity. Corneal stem cell dysfunction in diabetic patients as an important influencing factor of diabetic keratopathy. The consequences of this condition are often underestimated. The limbus is located between the cornea and the sclera tissue. The limbal stroma consists of a series of radial elevations with fibrovascular centers known as palisades of Vogt (POV). Previous studies have shown that palisades of Vogt (POV), as the main site of limbal stem cells, plays an important role in the homeostasis of the corneal epithelium. Therefore, POV plays a vital role in the healing of corneal epithelial surgery and postoperative evaluation. IVCM can observe the condition of the corneal epithelium at the cellular level. It has profound significance and guidance for the evaluation of limbal and limbal stem cells. We have previously observed structural changes in POV in HSK and HZO patients on IVCM. At present, there have been reports involving limbal stem cell dysfunction in diabetic patients, but the specific pathogenesis is still unclear. However, there are no studies on POV morphological changes in patients with DM. Therefore, we performed statistics and compared the correlation between POV morphological changes and corneal epithelial basal cell density, corneal nerves, and length of disease in DM patients and normal humans using IVCM studies. At the same time, fundoscopy was used to observe the correlation between the thickness of RNFL and the thickness of GCC and POV in diabetic patients. And to observe the correlation between SVD, DVD and POV for research.

Keywords: confocal microscopy, fundus, limbal stem cells, diabetes

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753 In vitro Modulation of Cytokine Expression by an Aqueous Licorice Extract in Canine

Authors: A. Watson, G. Telford, D. I. Pritchard

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Objective: We investigated the immunomodulatory ability of licorice (Glycyrrhiza glabra). Such activities could have value for the management of common immunological diseases in dogs, such as environmental allergy. This study investigated the potential of a Licorice root extract (LRE) to influence the relative expression of Th-1, Th-2, and Th-17 cytokines in canine peripheral blood mononuclear cells (PBMC). Methods: A LRE was prepared using an alcoholic-aqueous-based solvent method. The extract was tested in three in vitro assays using canine leukocytes to determine its toxicity and immunoregulatory profile. Extract toxicity was assessed using the human T-lymphocyte cell line, Jurkat E6.1. The impact of the extract on the proliferation of concanavalin-activated canine PBMC was also determined. Finally, the extract was assessed for its ability to influence cytokine release in activated PBMC, measuring culture medium concentrations of interleukin-17, interferon gamma, and interleukin-4. One-way ANOVA followed by Dunnett’s post-test was used for statistics using concanavalin positive control as reference (p ≤ 0.05). Results: There was evidence that the LRE had specific immunomodulatory properties, causing significant inhibition of IL4 expression over a non-toxic/non-cytostatic concentration range (p < 0.001). In the same cell incubations, there was no significant impact on IL17 nor IFNg over the same non-toxic/non-cytostatic concentration range. Conclusion: The study provides in vitro evidence that LRE preferentially reduces the expression of a Th-2-type cytokine, IL4. The dog population, as with humans, is prone to conditions associated with a Th-2 bias of the immune system, such as environmental allergy. Based on these results, licorice merits further evaluation as a useful immune modulator for such allergic diseases.

Keywords: cytokine, Glycyrrhiza glabra, peripheral blood mononuclear cells, T-cell activation

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752 Prospects of Milk Protein as a Potential Alternative of Natural Antibiotic

Authors: Syeda Fahria Hoque Mimmi

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Many new and promising treatments for reducing or diminishing the adverse effects of microorganisms are being discovered day by day. On the other hand, the dairy industry is accelerating the economic wheel of Bangladesh. Considering all these facts, new thoughts were developed to isolate milk proteins by the present experiment for opening up a new era of developing natural antibiotics from milk. Lactoferrin, an iron-binding glycoprotein with multifunctional properties, is crucial to strengthening the immune system and also useful for commercial applications. The protein’s iron-binding capacity makes it undoubtedly advantageous to immune system modulation and different bacterial strains. For fulfilling the purpose, 4 of raw and 17 of commercially available milk samples were collected from different farms and stores in Bangladesh (Dhaka, Chittagong, and Cox’s Bazar). Protein quantification by nanodrop technology has confirmed that raw milk samples have better quantities of protein than the commercial ones. All the samples were tested for their antimicrobial activity against 18 pathogens, where raw milk samples showed a higher percentage of antibacterial activity. In addition to this, SDS-PAGE (Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis) was performed to identify lactoferrin in the milk samples. Lactoferrin was detected in 9 samples from which 4 were raw milk samples. Interestingly, Streptococcus pyogenes, Klebsiella pneumoniae, Bacillus cereus, Pseudomonas aeruginosa, Vibrio cholera, Staphylococcus aureus, and enterotoxigenic E. coli significantly displayed sensitivity against lactoferrin collected from raw milk. Only Bacillus cereus, Pseudomonas aeruginosa, Streptococcus pneumonia, Enterococcus faecalis, and ETEC (Enterotoxigenic Escherichia coli) were susceptible to lactoferrin obtained from a commercial one. This study suggested that lactoferrin might be used as the potential alternative of antibiotics for many diseases and also can be used to reduce microbial deterioration in the food and feed industry.

Keywords: alternative of antibiotics, commercially available milk, lactoferrin, nanodrop technology, pathogens, raw milk

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751 Microbial and Oocyst Count in Feacal Material of Broilers Birds Administered Phytochemicals (Naringin and Hesperidin)

Authors: Adeleye Oluwagbemmiga, Obuotor Tolulope, Dosumu Adebisi, Opowoye I., Olasoju M., Kolawole Amos, Egbeyale Lawrence

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Gut Microbiota plays a vital role in animal health and welfare. This study investigated the effect of naringin and hesperidin administration on broiler birds. A total of 80 day – old broiler chicks were randomly divided into eight groups, with ten birds per group. Four groups were not inoculated but administered coccidiostat (1A), hesperidin alone (2A), naringin alone (3A) and a combination of naringin and hesperidin (4A) from day eight (8) to day fourteen (14) while four other groups (5A – 8A) were inoculated with 2 x 10⁴ oocysts per 0.5ml of Eimeria tenella on the 16th and 19th day of age after they were administered conventional antibiotics and coccidiostat, naringin (50mg/body weight), hesperidin (50mg/body weight) and a combination from day 8 - 14. McMaster counting technique was used to count the oocysts, while pour plate technique was used to determine the bacterial load. The results showed a significant increase in their performance with an average weight ranging from 1.55kg – 2.00kg, microbial load also improved with colony count values from 3.5 x 104 - 4.5 x 10⁴ CFU/ml. The study also found that the inclusion of naringin and hesperidin in the diets of broiler birds inoculated with coccidia oocysts significantly reduced the fecal oocyst counts, with the lowest count in combined treatment (8A) (10%) and indicating a lower degree of coccidiosis infection in the treated groups whereas control group (5A) had the highest oocyst count (35%). Mortality and Morbidity rate was 0% as none of the bird showed signs and symptoms. The reduction in oocyst counts could help to strengthen the immune system of broiler birds and limit the severity of coccidiosis infection, which could be an effective strategy for improving performance, immune function and mitigating the impact of coccidiosis infection in broiler birds.

Keywords: gut colonization, naringin, hesperidin, eimeria tenella, broilers

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750 Biocompatibility assessment of different origin Barrier Membranes for Guided Bone Regeneration

Authors: Antonio Munar-Frau, Sascha Klismoch, Manfred Schmolz, Federico Hernandez-Alfaro, Jordi Caballe-Serrano

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Introduction: Biocompatibility of biomaterials has been proposed as one of the main criteria for treatment success. For guided bone regeneration (GBR), barrier membranes present a conflict given the number of origins and modifications of these materials. The biologic response to biomaterials is orchestrated by a series of events leading to the integration or rejection of the biomaterial, posing questions such as if a longer occlusive property may trigger an inflammatory reaction. Whole blood cultures are a solution to study the immune response to drugs or biomaterials during the first 24-48 hours. The aim of this study is to determine the early immune response of different origins and chemical modifications of barrier membranes. Materials & Methods: 5 different widely used barrier membranes were included in this study: Acellular dermal matrix (AlloDerm, LifeCell®), Porcine Peritoneum (BioGide, Geistlich Pharma®), Porcine Pericardium (Jason, Botiss Biomaterials GmbH®), Porcine Cross-linked collagen (Ossix Plus, Datum Dental®) and d-PTFE (Cytoplast TXT, Osteogenics Biomedical®). Blood samples were extracted from 3 different healthy donors and incubated with the different samples of barrier membranes for 24 hours. After the incubation time, serum samples were obtained and analyzed by means of biocompatibility assays taking into account 42 markers. Results: In an early stage of the inflammatory response, the Acellular dermal matrix, porcine peritoneum and porcine cross-linked collagen expressed similar patterns of cytokine expression with a great manifestation of ENA 78. Porcine pericardium and d-PTFE presented similar cytokine activation, especially for MMP-3 and MMP-9, although other cytokines were highlighted with lower expression. For the later immune response, Porcine peritoneum and acellular dermal matrix MCP-1 and IL-15 were evident. Porcine pericardium, porcine cross-linked collagen and d-PTFE presented a high expression of IL-16 and lower manifestation of other cytokines. Different behaviors depending on an earlier or later stage of the inflammation process were observed. Barrier membrane inflammatory expression does not only differ depending on the origin, variables such as treatment of the collagen and polymers may also have a great impact on the cytokine expression of the studied barrier membranes during inflammation. Conclusions: Surface treatment and modifications might affect the biocompatibility of the membranes, as different cytokine expressions were evidently depending on the origin of the biomaterial. This study is only a brushstroke regarding the biocompatibility of materials, as it is one of the pioneer studies for ex vivo barrier membranes assays. Studies regarding surface modification are needed in order to clarify mystifications of barrier membrane science.

Keywords: biomaterials, bone regeneration, biocompatibility, inflammation

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749 Laboratory-Based Monitoring of Hepatitis B Virus Vaccination Status in North Central Nigeria

Authors: Nwadioha Samuel Iheanacho, Abah Paul, Odimayo Simidele Michael

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Background: The World Health Assembly through the Global Health Sector Strategy on viral hepatitis calls for the elimination of viral hepatitis as a public health threat by 2030. All hands are on deck to actualize this goal through an effective and active vaccination and monitoring tool. Aim: To combine the Epidemiologic with Laboratory Hepatitis B Virus vaccination monitoring tools. Method: Laboratory results analysis of subjects recruited during the World Hepatitis week from July 2020 to July 2021 was done after obtaining their epidemiologic data on Hepatitis B virus risk factors, in the Medical Microbiology Laboratory of Benue State University Teaching Hospital, Nigeria. Result: A total of 500 subjects comprising males 60.0%(n=300/500) and females 40.0%(n=200/500) were recruited. A fifty-three percent majority was of the age range of 26 to 36 years. Serologic profiles were as follows, 15.0%(n=75/500) HBsAg; 7.0% (n=35/500) HBeAg; 8.0% (n=40/500) Anti-Hbe; 20.0% (n=100/500) Anti-HBc and 38.0% (n=190/500) Anti-HBs. Immune responses to vaccination were as follows, 47.0%(n=235/500) Immune naïve {no serologic marker + normal ALT}; 33%(n=165/500) Immunity by vaccination {Anti-HBs + normal ALT}; 5%(n=25/500) Immunity to previous infection {Anti-HBs, Anti-HBc, +/- Anti-HBe + normal ALT}; 8%(n=40/500) Carriers {HBsAg, Anti-HBc, Anti-HBe +normal ALT} and 7% (35/500) Anti-HBe serum- negative infections {HBsAg, HBeAg, Anti-HBc +elevated ALT}. Conclusion: The present 33.0% immunity by vaccination coverage in Central Nigeria was much lower than the 41.0% national peak in 2013, and a far cry from the global expectation of attainment of a Global Health Sector Strategy on the elimination of viral hepatitis as a public health threat by 2030. Therefore, more creative ideas and collective effort are needed to attain this goal of the World Health Assembly.

Keywords: Hepatitis B, vaccination status, laboratory tools, resource-limited settings

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