Search results for: immune cell

Authors: Bit-Na Kwon, Hyun-Jee Yang, Dong-Hyun Ha, Hyoung-Kyu Song

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In this paper, an adaptive cooperative scheme in the cell edge is proposed. The proposed scheme considers the location of a user and applies the suitable cooperative scheme. In cellular systems, the performance of communication is degraded if the user is located in the cell edge. In conventional scheme, two base stations are used in order to obtain diversity gain. However, the performance of communication is not sufficiently improved since the distance between each base station and a user is still distant. Therefore, we propose a scheme that the relays are used and the cooperative scheme is adaptively applied according to the user location. Through simulation results, it is confirmed that the proposed scheme has better performance than the conventional scheme.

Keywords: adaptive transmission, cooperative communication, diversity gain, OFDM

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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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Authors: Pratiksha Dubey, Praveen Singh, Shantanu Sen Gupta, Anand K. Bachhawat

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5-oxoproline (pyroglutamic acid) is a cyclic lactam of glutamic acid. In the cell, it can be produced by several different pathways and is metabolized into glutamate with the help of the 5-oxoprolinase enzyme (OPLAH or OXP1). The inhibition of 5-oxoprolinase enzyme in mammals was found to result in heart failure and is thought to be a consequence of oxidative stress [1]. To analyze the consequences of 5-oxoproline accumulation more clearly, we are generating models for 5-oxoproline accumulation in yeast. The 5-oxoproline accumulation model in yeast is being developed by two different strategies. The first one is by overexpression of the mouse  -glutamylcyclotransferase enzyme. It degrades -glu-met dipeptide into 5-oxoproline and methionine taken by the cell from the medium. The second strategy is by providing high concentration of 5-oxoproline externally to the yeast cells. The intracellular 5-oxoproline levels in both models are being evaluated. In addition, the metabolic and cellular consequences are being investigated.

Keywords: 5-oxoproline, pyroglutamic acid, yeast, genetics

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Authors: Martin Van, Mohammed Javed, Sarah Hemington-Gorse

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Aim: Our aim was to determine the effect of size and tumor depth of basal cell carcinomas (BCCs) on surgical margin clearance. Methods: A retrospective study was conducted at the Welsh Centre for Burns and Plastic Surgery (WCBPS), Morriston Hospital between 1 Jan 2016 – 31 July 2016. Only patients with confirmed BCC on histopathological analysis were included. Patient data including anatomical region treated, lesion size, histopathological clearance margins and histological sub-types were recorded. An independent T-test was performed determine statistical significance. Results: A total of 228 BCCs were excised in 160 patients. Eleven lesions (4.8%) were incompletely excised. The nose area had the highest rate of incomplete excision. The mean diameter of incompletely excised lesions was 11.4mm vs 11.5mm in completely excised lesions (p=0.959) and the mean histological depth of incompletely excised lesions was 4.1mm vs. 2.5mm for completely excised BCCs (p < 0.05). Conclusions: BCC tumor depth of > 4.1 mm was associated with high rate of incomplete margin clearance. Hence, in prospective patients, a BCC tumor depth (>4 mm) on tissue biopsy should alert the surgeon of potentially higher risk of incomplete excision of lesion.

Keywords: basal cell carcinoma, excision margins, plastic surgery, treatment

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Authors: Srisopa Ruangnoo, Arunporn Itharat

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The ethanolic and aqueous extracts of Parkia javanica Merr. germinating seeds and Parkia speciosa Hassk. seeds were evaluated for cytotoxic activity against three different types of human cancer cell lines including colon cancer (LS174T), breast cancer (MCF-7) and prostate cancer (PC3) using sulforhodamine B (SRB) assay. The fresh plant parts were divided into 2 parts. The first part was extracted by maceration with 95% ethanol for 3 days and then filtered, and the filtrates were evaporated by rotary evaporator. The other part was squeezed and filtered. Then the filtrates were dried by freeze dryer. The screening found that the aqueous extract of P. javanica Merr. germinating seeds exhibited more than 70% inhibition (at concentration 50 µg/ml) against all types of human cancer cells. The aqueous extract of P. javanica Merr. germinating seeds showed the highest cytotoxic activity against MCF-7 with the IC50 value as 5.63 µg/ml. The aqueous extract of P. javanica Merr. germinating seeds also showed high cytotoxic activity against PC3 and LS174T with the IC50 values as 10.79 and 11.40 µg/ml, respectively. In conclusion, P. javanica Merr. germinating seed is a natural source of anticancer activity and further research to isolate active compounds from this plant should be undertaken.

Keywords: cytotoxic activity, Parkia javanica Merr., Parkia speciosa Hassk., human cancer cell lines

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Authors: Svetlana Guryeva, Inna Kornienko, Andrey Usanov, Dmitry Usanov, Elena Petersen

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Every day cells in our organism are exposed to various factors: chemical agents, reactive oxygen species, ionizing radiation, and others. These factors can cause damage to DNA, cellular membrane, intracellular compartments, and proteins. The fate of cells depends on the exposure intensity and duration. The prolonged and intense exposure causes the irreversible damage accumulation, which triggers the permanent cell cycle arrest (cellular senescence) or cell death programs. In the case of low dose of impacts, it can lead to cell renovation and to cell functional state improvement. Therefore, it is a pivotal question to investigate the factors and doses that result in described positive effects. In order to estimate the influence of different agents, the proliferation index and levels of cell death markers (annexin V/propidium iodide), senescence-associated β-galactosidase, and lipofuscin were measured. The experiments were conducted on primary human fibroblasts of the 8th passage. According to the levels of mentioned markers, these cells were defined as senescent cells. The effect of low-frequency magnetic field was investigated. Different modes of magnetic field exposure were tested. The physical agents were compared with chemical agents: metformin (10 mM) and taurine (0.8 mM and 1.6 mM). Cells were incubating with chemicals for 5 days. The highest decrease in the level of senescence-associated β-galactosidase (21%) and lipofuscin (17%) was observed in the primary senescent fibroblasts after 5 days after double treatments with 48 h intervals with low-frequency magnetic field. There were no significant changes in the proliferation index after magnetic field application. The cytotoxic effect of magnetic field was not observed. The chemical agent taurine (1.6 mM) decreased the level of senescence-associated β-galactosidase (23%) and lipofuscin (22%). Metformin improved the activity of senescence-associated β-galactosidase on 15% and the level of lipofuscin on 19% in this experiment. According to these results, the effect of double treatment with 48 h interval with low-frequency magnetic field and the effect of taurine (1.6 mM) were comparable to the effect of metformin, for which anti-aging properties are proved. In conclusion, this study can become the first step towards creation of the standardized system for the investigation of different effects on senescent cells.

Keywords: biomarkers, magnetic field, metformin, primary fibroblasts, senescence, taurine

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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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Authors: Sun Il Yu, Jung Hyun Joo, Hwa Sung Shin

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Astrocytes are known as dominant cells in CNS and play a role as a supporter of CNS activity and regeneration. Recently, three-dimensional culture of astrocytes were actively applied to understand in vivo astrocyte works. Electrospun nanofibers are attractive for 3D cell culture system because they have a high surface to volume ratio and porous structure, and have already been used for 3D astrocyte cultures. In this research, the stiffness of cellulose acetate (CA) nanofiber was controlled by heat treatment. As stiffness increased, astrocyte cell viability and adhesion increased. Reactivity of astrocyte was also upregulated in stiffer CA nanofiber in terms of GFAP, an intermediate filament protein. Finally, we demonstrated that stiffness-controllable CA is attractive for astrocyte tissue engineering.

Keywords: astrocyte, cellulose acetate, nanofiber, tissue scaffold

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Authors: Ashok K. Gupta

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Over the past few decades, since the bio-engineering revolution, autologous cell therapy (ACT) has become a rapidly evolving field. Currently, this form of therapy has broad applications in modern medicine and plastic surgery, ranging from the treatment/improvement of wound healing to life-saving operations. A study was conducted on 50 patients having to disfigure, and deform post burn scars and was treated by injection of extracted, refined adipose tissue grafts with their unique stem cell properties. To compare the outcome, a control of 20 such patients was treated with conventional skin or soft-tissue flaps or skin grafting, and a control of 10 was treated with more advanced microsurgical techniques such as Pre-fabricated flaps/pre laminated flaps / free flaps. Assessment of fat volume and survival post- follow up period was done by radiological aid, using MRI and clinically (Survival of the autograft and objective parameters for scar elasticity were evaluated skin elasticity parameters 3 to 9 months postoperatively). Recently, an enzyme that is involved in collagen crosslinking in fibrotic tissue, lysyl hydroxylase (LH2), was identified. This enzyme is normally active in bone and cartilage but hardly in the skin. It has been found that this enzyme is highly expressed in scar tissue and subcutaneous fat; this is in contrast to the dermis, where the enzyme is hardly expressed. Adipose tissue-derived stem cell injections are an effective method in the treatment of various extensive post-burn scar deformities that makes it possible to re-create the lost sub-dermal tissue for improvement in the function of involved joint movements.

Keywords: adipose tissue-derived stem cell injections, treatment of various extensive post-burn scar deformities, re-create the lost sub-dermal tissue, improvement in function of involved joint movements

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Authors: Azeez Shaik, Yagnik Kalariya, Amit Salvi

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High-performance requirements and emission norms were forcing the automobile industry to opt for lightweight materials which improve the fuel efficiency and absorb energy during crash applications. In such scenario, the woven fabric composites are providing better energy absorption compared to metals. Woven fabric composites have a repetitive unit cell (RUC) and the mechanical properties of these materials are highly dependent on RUC. This work investigates the importance of detailed modelling of the RUC, the size effects associated and the mitigation techniques to avoid them using Finite element analysis approach.

Keywords: repetitive unit cell, representative volume element, size effects, cohesive zone, finite element analysis

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Authors: Melliti Rihab, Zaeid Sonia, Khechine Wiem, Daldoul Amira

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Introduction: Lung cancer is the leading cause of cancer death. Its incidence is increasing, and its prognosis remains pejorative. We present the clinical, pathological, and therapeutic characteristics of bronchopulmonary cancer (BPC) in Tunisia. Methods: Retrospective study including patients followed in the oncology department of the University Hospital of Monastir between April 2014 and December 2021 suffering from lung cancer. Results: These are 117 patients, including 86.3% men and 13.7% women (sex ratio 6.3). The average age was 64 years ± 9 (37-83), with 95.7% being over 50 years old. Patients were smokers in 82% of cases. The clinical signs were dominated by chest pain (27.5%) and dyspnea in 21.1% of cases. In 6 patients, an episode of COVID-19 infection revealed the diagnosis. Half of the patients had a PS between 0 and 1. Small cell lung cancer was present in 18 patients (15.4%). The majority of non small cell lung cancer was of the adenocarcinoma type (68.7%). The diagnosis was late (stage IV) in 62.4% of cases. BPC was metastatic to bone (52%), contralateral lung (25.9%), and brain (27.3%). Patients were oligometastatic in 26% of cases. Surgery and radiotherapy were performed respectively in 14.5% and 23.1% of cases. Three-quarters of the patients had had nutrition (75.2%). The ROS1 mutation was present in 1 patient. PDL-1 expression was >40% in 2 patients. Survival was mean eight months ± 7.4. Conclusion: Lung cancer is diagnosed at a late stage in Tunisia. The lack of molecular study for non-small cell PBC and the lack of marketing authorization for tyrosine kinase inhibitors in Tunisia make the management incomplete.

Keywords: SCLC, NCSLC, ROS1, PDL1

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Authors: Sara Kamalisiahroudi, Zhang Jianbo, Bin Wu, Jun Huang, Laisuo Su

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The temperature rising within a battery cell depends on the level of heat generation, the thermal properties and the heat transfer around the cell. The rising of temperature is a serious problem of Lithium-Ion batteries and the internal resistance of battery is the main reason for this heating up, so the heat generation rate of the batteries is an important investigating factor in battery pack design. The delivered power of a battery is directly related to its capacity, decreases in the battery capacity means the growth of the Solid Electrolyte Interface (SEI) layer which is because of the deposits of lithium from the electrolyte to form SEI layer that increases the internal resistance of the battery. In this study two identical cylindrical Lithium-Ion (NCR18650)batteries from the same company with noticeable different in capacity (a fresh and a used battery) were compared for more focusing on their heat generation parameters (entropy coefficient and internal resistance) according to Brandi model, by utilizing potentiometric method for entropy coefficient and EIS method for internal resistance measurement. The results clarify the effect of capacity difference on cell electrical (R) and thermal (dU/dT) parameters. It can be very noticeable in battery pack design for its Safety.

Keywords: heat generation, Solid Electrolyte Interface (SEI), potentiometric method, entropy coefficient

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Authors: Yen-Ni Teng, Hsing-Yi Chen, Hsien-An Pan, Yung-Ming Lin, Hany A. Omar, Jui-Hsiang Hung

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Leucine-rich repeats and WD repeat domain containing 1 (LRWD1) is highly expressed in the testes of healthy males. On the other hand, LRWD1 is significantly down-regulated in the testicular tissues of patients with severe spermatogenic defects. In our study, the downregulation of LRWD1 expression by shRNA caused a significant reduction of cell growth and mitosis and a noteworthy increase in the cell microtubule atrophy rate. Here, we used EMBOSS CpG plot analysis to explore the promoter region of LRWD1 gene. We found that CpG islands are located between positions -253 to +5 nucleotides upstream from the LRWD1 transcription start site. Luciferase reporter assay revealed that the hypermethylation of the LRWD1 promoter reduced the transcription activity in cells. In addition, quantitative methylation-specific PCR and immunostaining showed that the methylation inhibitor, 5-Aza-2'-deoxycytidine, increased LRWD1 promoter activity, LRWD1 mRNA, protein expression and cell viability. Whereas, the methylation activator, S-adenosylmethionine, caused opposite effects. The overexpression of p53 and Nrf2 in NT2/D1 cells increased LRWD1 promoter activity while 5-fluorodeoxyuridine decreased it. In conclusion, this study highlights evidence that the methylation status of LRWD1 promoter is associated with LRWD1 expression. Since the expression level of LRWD1 plays an important role in spermatogenesis, the methylation status of LRWD1 may serve as a novel molecular diagnostic or therapeutic approach in male's infertility.

Keywords: LRWD1, DNA methylation, p53, Nrf2

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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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Authors: Gourav Jain, Sameer Dixit, Surjeet Kumar Arya, Praveen C. Verma

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Plant cell wall plays a major role in defence mechanism against biotic and abiotic stress as it constitutes the physical barrier between the microenvironment and internal component of the cell. It is a complex structure composed of mostly carbohydrates among which cellulose and hemicelluloses are most abundant that is embedded in a matrix of pectins and proteins. Multiple enzymes have been reported which plays a vital role in cell wall modification, Pectin Methylesterase (PME) is one of them which catalyses the demethylesterification of homogalacturonans component of pectin which releases acidic pectin and methanol. As emitted methanol is toxic to the insect pest, we use PME gene for the better methanol production. In the current study we showed overexpression of PME gene isolated from Withania somnifera under the insect inducible promoter causes enhancement of methanol production at the time of insect feeds to plants, and that provides better insect resistance property. We found that the 85-90% mortality causes by transgenic tobacco in both chewing (Spodoptera litura larvae and Helicoverpa armigera) and sap-sucking (Aphid, mealybug, and whitefly) pest. The methanol content and emission level were also enhanced by 10-15 folds at different inducible time point interval (15min, 30min, 45min, 60min) which would be analysed by Purpald/Alcohol Oxidase method.

Keywords: methanol, Pectin methylesterase, inducible promoters, Purpald/Alcohol oxidase

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Authors: Majid Moshirfar, Kyle Margulies, Yasmyne C. Ronquillo, Phillip Hoopes

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A total of 66 articles were reviewed to compare glaucoma and its associated surgeries' effect on central corneal endothelium cell density (CECD). The paper reports the average reported central CECD loss at 3-, 6-, 12-, 24-, 36-, 48-, and 60-month post-operation for each glaucoma surgery. ALT, MLT, SLT, CS AGV, VC BGI, Hydrus + phaco, XEN gel + phaco, PRESERFLO, Dual iStent, or Trabectome had no significant impact on postoperative CECD compared to either preoperative CECD or control group CECD. The highest CECD loss was found to be EXPRESS-phaco, AC AGV, CS BGI, CS BGI, AC BGI, and AC BGI at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups, respectively. AC AGV, Trab + MMC, Trab, AC BGI, Trab + MMC, Cypass, and Cypass showed the smallest reduction of CECD at the 3-, 6-, 12-, 24-, 36-, 48-, and 60-month follow-ups.

Keywords: glaucoma, corneal endothelium, cell density, surgery outcome

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Authors: Urszula Stachewicz

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Surface properties of materials are the key parameter in many applications, especially in the biomedical field, to control cell-material interactions. In our work, we want to achieve the controllability of surface properties of polymer fibers via a single-step electrospinning process by alternating voltage polarities. Voltage polarity defines the charge accumulated on the surface of the liquid jet and the surface of the fibers. Positive polarity attracts negatively charged groups to fibers’ surface, whereas negative polarity moves the negatively charged functional groups away from the surface. This way, we can control the surface chemistry, wettability, and additionally surface potential of electrospun fibers. Within our research, we characterized surface chemistry using X-ray photoelectron microscopy (XPS) and surface potential with Kelvin probe force microscopy (KPFM) on electrospun fibers of commonly used polymers such as PCL, PVDF, and PMMA, often used as biomaterials. We proved the significant effect of fibers' surface potential on cell integration with the scaffolds and further cells development for the regeneration processes based on the osteoblast and fibroblast culture studies. Acknowledgments: The study was conducted within ‘Nanofiber-based sponges for atopic skin treatment’ project, which is carried out within the First TEAM programme of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund, project no POIR.04.04.00-00- 4571/18-00.

Keywords: cell attachment, fibers, fibroblasts, osteoblast, proliferation, surface potential

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Authors: Sima Parvizi Omran, Rezvan Tavakoli, Mahnaz Safari, Mohammadreza Aghasadeghi, Abolfazl Fateh, Pooneh Rahimi

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Background: SARS-CoV-2, a single-stranded RNA betacoronavirus causes the global outbreak of coronavirus disease 2019 (COVID-19). Several clinical and scientific concerns are raised by this pandemic. Genetic factors can contribute to pathogenesis and disease susceptibility. There are single nucleotide polymorphisms (SNPs) in many of the genes in the immune system that affect the expression of specific genes or functions of some proteins related to immune responses against viral infections. In this study, we analyzed the impact of polymorphism in the interferon alpha and beta receptor subunit 2 (IFNAR2) and dipeptidyl peptidase 9 (Dpp9) genes and clinical parameters on the susceptibility and resistance to Coronavirus disease (COVID-19). Methods: A total of 330- SARS-CoV-2 positive patients (188 survivors and 142 nonsurvivors) were included in this study. All single-nucleotide polymorphisms (SNPs) on IFNAR2 (rs2236757) and Dpp9 (rs2109069) were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Results: In survivor patients, the frequency of the favourable genotypes of IFNAR2 SNP (rs2236757 GC) was significantly higher than in nonsurvivor patients, and also Dpp9 (rs2109069 AT) genotypes were associated with the severity of COVID-19 infection. Conclusions: This study demonstrated that the severity of COVID- 19 patients was strongly associated with clinical parameters and unfavourable IFNAR2, Dpp9 SNP genotypes. In order to establish the relationship between host genetic factors and the severity of COVID-19 infection, further studies are needed in multiple parts of the world.

Keywords: SARS-CoV-2, COVID-19, interferon alpha and beta receptor subunit 2, dipeptidyl peptidase 9, single-nucleotide polymorphisms

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Authors: G. A. Amin, A. D. Al-Talhi

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The bacterium B. subtilis produced surfactin in conventional batch culture as a growth associated product and a growth rate (0.4 h-1). A fed-batch process was developed and the fermentative substrate and other nutrients were fed on hourly basis and according to the growth rate of the bacterium. Conversion of different quantities of Maldex-15 into surfactin was investigated in five different fermentation runs. In all runs, most of Maldex-15 was consumed and converted into surfactin and cell biomass with appreciable efficiencies. The best results were obtained with fermentation run supplied with 200 g Maldex-15. Up to 35.4 g.l-1 of surfactin and cell biomass of 30.2 g.l-1 were achieved in 12 hrs. Also, markedly substrate yield of 0.269 g/g and volumetric reactor productivity of 2.61 g.1-1.h-1 were obtained confirming the establishment of a cost effective commercial surfactin production.

Keywords: Bacillus subtilis, biosurfactant, exponentially fed-batch fermentation, surfactin

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Authors: S. Vasailor, C. Rattanakawin

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Electro-winning of copper metal from dilute sulfate solution (13.7 g/L) was performed in a lab electrolytic cell with stainless-steel cathode and lead-alloy anode. The effects of various parameters including cell voltage, electro-winning temperature and time were studied in order to acquire an appropriate current efficiency of copper deposition. The highest efficiency is about 95% obtaining from electro-winning condition of 3V, 55°C and 3,600 s correspondingly. The cathode copper with 95.5% Cu analyzed using atomic absorption spectrometry can be obtained from this single-winning condition. In order to increase the copper grade, solvent extraction should be used to increase the sulfate concentration, say 50 g/L, prior to winning the cathode copper effectively.

Keywords: copper metal, current efficiency, dilute sulfate solution, electro-winning

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Authors: Ateeqa Aslam, Hans J. Nauwynck

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Porcine hemagglutinating encephalomyelitis virus (PHEV) is a betacoronavirus that has been studied in the past as a cause of vomiting and wasting disease (VWD) in young piglets (<3 weeks). Nowadays, the virus is still circulating on most farms in Belgium, but there are no descriptions anymore of VWD. Therefore, we are interested in differences between the old and new strains. We compared the replication kinetics of the old well-studied strain VW572 (1972) and the recent isolate P412 (2020) in a susceptible continuous cell line (RPD cells) and in primary porcine respiratory epithelial cells (PoRECs). The RPD cell line was inoculated with each PHEV strain at an m.o.i. of 1 the supernatant was collected, and the cells were fixed at different time points post-inoculation. The supernatant was titrated (extracellular virus titer), and the infected cells were revealed by immunofluorescence staining and quantitated by fluorescence microscopy. We found that VW572 replicated better in the RPD cell line at earlier time points when compared to P412. Porcine respiratory epithelial cells (PoREC) were isolated, grown at air-liquid interphase in transwells and inoculated with both strains of PHEV at a virus titer of 106.6TCID50 per 200 µl either at the apical side or at the basal side of the cells. At different time points after inoculation, the transwells were fixed and stained for infected cells. VW572 preferentially infected the epithelial cells via the basolateral side of porcine nasal epithelial cells, whereas P412 preferred the apical side. These findings suggest that there has been an evolution of PHEV in its interaction with the respiratory epithelial cells. In the future, more virus strains will be enclosed and the tropism of the strains for different neuronal cell types will be examined for the change in virus neurotropism.

Keywords: porcine hemagglutinating encephalomyelitis virus (PHEV), primary porcine respiratory epithelial cells (PoRECs), virus tropism, vomiting and wasting disease (VWD)

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Authors: Dugeree Otgongerel, Kyong Jin Cho, Yu-Han Kim, Sangmee Ahn Jo

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Excessive activation of glutamate receptor is thought to be involved in retinal ganglion cell (RGC) death after ischemia- reperfusion damage. Glutamate carboxypeptidase II (GCPII) is an enzyme responsible for the synthesis of glutamate. Several studies showed that inhibition of GCPII prevents or reduces cellular damage in brain diseases. Thus, in this study, we examined the expression of GCPII in rat retina and the role of GCPII in acute high IOP ischemia-reperfusion damage of eye by using a GCPII inhibitor, 2-(phosphonomethyl) pentanedioic acid (2-PMPA). Animal model of ischemia-reperfusion was induced by raising the intraocular pressure for 60 min and followed by reperfusion for 3 days. Rats were randomly divided into four groups: either intra-vitreous injection of 2-PMPA (11 or 110 ng per eye) or PBS after ischemia-reperfusion, 2-PMPA treatment without ischemia-reperfusion and sham-operated normal control. GCPII immunoreactivity in normal rat retina was detected weakly in retinal nerve fiber layer (RNFL) and retinal ganglionic cell layer (RGL) and also inner plexiform layer (IPL) and outer plexiform layer (OPL) strongly where are co-stained with an anti-GFAP antibody, suggesting that GCPII is expressed mostly in Muller and astrocytes. Immunostaining with anti-BRN antibody showed that ischemia- reperfusion caused RGC death (31.5 %) and decreased retinal thickness in all layers of damaged retina, but the treatment of 2-PMPA twice at 0 and 48 hour after reperfusion blocked these retinal damages. GCPII level in RNFL layer was enhanced after ischemia-reperfusion but was blocked by PMPA treatment. This result was confirmed by western blot analysis showing that the level of GCPII protein after ischemia- reperfusion increased by 2.2- fold compared to control, but this increase was blocked almost completely by 110 ng PMPA treatment. Interestingly, GFAP immunoreactivity in the retina after ischemia- reperfusion followed by treatment with PMPA showed similar pattern to GCPII, increase after ischemia-reperfusion but reduction to the normal level by PMPA treatment. Our data demonstrate that increase of GCPII protein level after ischemia-reperfusion injury is likely to cause glial activation and/or retinal cell death which are mediated by glutamate, and GCPII inhibitors may be useful in treatment of retinal disorders in which glutamate excitotoxicity is pathogenic.

Keywords: glutamate carboxypepptidase II, glutamate excitotoxicity, ischemia-reperfusion, retinal ganglion cell

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Authors: I. Székács, Á. Fejes, S. Klátyik, E. Takács, D. Patkó, J. Pomóthy, M. Mörtl, R. Horváth, E. Madarász, B. Darvas, A. Székács

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Environmental and toxicological characteristics of formulated pesticides may substantially differ from those of their active ingredients or other components alone. This phenomenon is demonstrated in the case of the herbicide active ingredient glyphosate. Due to its extensive application, this active ingredient was found in surface and ground water samples collected in Békés County, Hungary, in the concentration range of 0.54–0.98 ng/ml. The occurrence of glyphosate appeared to be somewhat higher at areas under intensive agriculture, industrial activities and public road services, but the compound was detected at areas under organic (ecological) farming or natural grasslands, indicating environmental mobility. Increased toxicity of the formulated herbicide product Roundup, compared to that of glyphosate was observed on the indicator aquatic organism Daphnia magna Straus. Acute LC50 values of Roundup and its formulating adjuvant Polyethoxylated Tallowamine (POEA) exceeded 20 and 3.1 mg/ml, respectively, while that of glyphosate (as isopropyl salt) was found to be substantially lower (690-900 mg/ml) showing good agreement with literature data. Cytotoxicity of Roundup, POEA and glyphosate has been determined on the neuroectodermal cell line, NE-4C measured both by cell viability test and holographic microscopy. Acute toxicity (LC50) of Roundup, POEA and glyphosate on NE-4C cells was found to be 0.013±0.002%, 0.017±0.009% and 6.46±2.25%, respectively (in equivalents of diluted Roundup solution), corresponding to 0.022±0.003 and 53.1±18.5 mg/ml for POEA and glyphosate, respectively, indicating no statistical difference between Roundup and POEA and 2.5 orders of magnitude difference between these and glyphosate. The same order of cellular toxicity seen in average cell area has been indicated under quantitative cell visualization. The results indicate that toxicity of the formulated herbicide is caused by the formulating agent, but in some parameters toxicological synergy occurs between POEA and glyphosate.

Keywords: glyphosate, polyethoxylated tallowamine, Roundup, combined aquatic and cellular toxicity, synergy

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Authors: Z. Nurzaireena, K. Azalea, T. Azirawaty, S. Jameela, G. Muralitharan

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The aim of this study is the review of the management practices of sickle cell disease patients during pregnancy, as well as the maternal and neonatal outcome at Ampang Hospital, Selangor. The study consisted of a review of pregnant patients with sickle cell disease under follow up at the Hematology Clinic, Ampang Hospital over the last seven years to assess their management and maternal-fetal outcome. The results of the review show that Ampang Hospital is considered the public hematology centre for sickle cell disease and had successfully managed three pregnancies throughout the last seven years. Patients’ presentations, managements and maternal-fetal outcome were compared and reviewed for academic improvements. All three patients were seen very early in their pregnancy and had been given a regime of folic acid, antibiotics and thrombo-prophylactic drugs. Close monitoring of maternal and fetal well being was done by the hematologists and obstetricians. Among the patients, there were multiple admissions during the pregnancy for either a painful sickle cell bone crisis, haemolysis following an infection and anemia requiring phenotype- matched blood and exchange transfusions. Broad spectrum antibiotics coverage during and infection, hydration, pain management and venous-thrombolism prophylaxis were mandatory. The pregnancies managed to reach near term in the third trimester but all required emergency caesarean section for obstetric indications. All pregnancies resulted in live births with good fetal outcome. During post partum all were nursed closely in the high dependency units for further complications and were discharged well. Post partum follow up and contraception counseling was comprehensively given for future pregnancies. Sickle cell disease is uncommonly seen in the East, especially in the South East Asian region, yet more cases are seen in the current decade due to improved medical expertise and advance medical laboratory technologies. Pregnancy itself is a risk factor for sickle cell patients as increased thrombosis event and risk of infections can lead to multiple crisis, haemolysis, anemia and vaso-occlusive complications including eclampsia, cerebrovasular accidents and acute bone pain. Patients mostly require multiple blood product transfusions thus phenotype-matched blood is required to reduce the risk of alloimmunozation. Emphasizing the risks and complications in preconception counseling and establishing an ultimate pregnancy plan would probably reduce the risk of morbidity and mortality to the mother and unborn child. Early management for risk of infection, thromboembolic events and adequate hydration is mandatory. A holistic approach involving multidisciplinary team care between the hematologist, obstetricians, anesthetist, neonatologist and close nursing care for both mother and baby would ensure the best outcome. In conclusion, sickle cell disease by itself is a high risk medical condition and pregnancy would further amplify the risk. Thus, close monitoring with combine multidisciplinary care, counseling and educating the patients are crucial in achieving the safe outcome.

Keywords: anaemia, haemoglobinopathies, pregnancy, sickle cell disease

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Authors: Sina Mahdavi, Mahdi Asghari Ozma

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Multiple sclerosis (MS) is the most common inflammatory autoimmune disease of the CNS that affects the myelination process in the central nervous system (CNS). Complex interactions of various "environmental or infectious" factors may act as triggers in autoimmunity and disease progression. The association between viral infections, especially human Varicella-zoster virus (VZV) and MS is one potential cause that is not well understood. This study aims to summarize the available data on VZV retrovirus infection in MS disease progression. For this study, the keywords "Multiple sclerosis", " Human Varicella-zoster virus ", and "central nervous system" in the databases PubMed, Google Scholar, Sid, and MagIran between 2016 and 2022 were searched and 14 articles were chosen, studied, and analyzed. Analysis of the amino acid sequences of HNRNPA1 with VZV proteins has shown a 62% amino acid sequence similarity between VZV gE and the PrLD/M9 epitope region (TNPO1 binding domain) of mutant HNRNPA1. A heterogeneous nuclear ribonucleoprotein (hnRNP), which is produced by HNRNPA1, is involved in the processing and transfer of mRNA and pre-mRNA. Mutant HNRNPA1 mimics gE of VZV as an antigen that leads to autoantibody production. Mutant HnRNPA1 translocates to the cytoplasm, after aggregation is presented by MHC class I, followed by CD8 + cells. Of these, antibodies and immune cells against the gE epitopes of VZV remain due to the memory immune response, causing neurodegeneration and the development of MS in genetically predisposed individuals. VZV expression during the course of MS is present in genetically predisposed individuals with HNRNPA1 mutation, suggesting a link between VZV and MS, and that this virus may play a role in the development of MS by inducing an inflammatory state. Therefore, measures to modulate VZV expression may be effective in reducing inflammatory processes in demyelinated areas of MS patients in genetically predisposed individuals.

Keywords: multiple sclerosis, varicella-zoster virus, central nervous system, autoimmunity

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Authors: H. Fuad Usman, M. Rafay Khan Sial, Shahzaib Hamid

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The trend of renewable energy resources has been amplified due to global warming and other environmental related complications in the 21st century. Recent research has very much emphasized on the generation of electrical power through renewable resources like solar, wind, hydro, geothermal, etc. The use of the photovoltaic cell has become very public as it is very useful for the domestic and commercial purpose overall the world. Although a single cell gives the low voltage output but connecting a number of cells in a series formed a complete module of the photovoltaic cells, it is becoming a financial investment as the use of it fetching popular. This also reduced the prices of the photovoltaic cell which gives the customers a confident of using this source for their electrical use. Photovoltaic cell gives the MPPT at single specific point of operation at a given temperature and level of solar intensity received at a given surface whereas the focal point changes over a large range depending upon the manufacturing factor, temperature conditions, intensity for insolation, instantaneous conditions for shading and aging factor for the photovoltaic cells. Two improved algorithms have been proposed in this article for the MPPT. The widely used algorithms are the ‘Incremental Conductance’ and ‘Perturb and Observe’ algorithms. To extract the maximum power from the source to the load, the duty cycle of the convertor will be effectively controlled. After assessing the previous techniques, this paper presents the improved and reformed idea of harvesting maximum power point from the photovoltaic cells. A thoroughly go through of the previous ideas has been observed before constructing the improvement in the traditional technique of MPP. Each technique has its own importance and boundaries at various weather conditions. An improved technique of implementing the use of both ‘Perturb and Observe’ and ‘Incremental Conductance’ is introduced.

Keywords: duty cycle, MPPT (Maximum Power Point Tracking), perturb and observe (P&O), photovoltaic module

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Authors: Ghada Esheba, Ebtisam Aljerayan, Afnan Al-Ghamdi, Atheer Alsharif, Hanan alzahrani

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Background: Primary ovarian neoplasms comprise a heterogeneous group of tumors of three main subtypes: surface epithelial, germ cell, and sex cord-stromal. The wide morphological variation within and between these groups can result in diagnostic difficulties. Gonadal sex cord-stromal tumors (SCST) represent one of the most heterogeneous categories of human neoplasms, because they may contain various combinations of different gonadal sex cord and stromal element. Aim: The aim of this work is to highlight the clinicopathological characteristics of SCST and to assess the value of alpha-inhibin and calretinin in the distinction between SCST and their mimics. Material and methods: This study was carried out on 100 cases using full tissue sections; 70 cases were SCST and 30 cases were histological mimics of SCST. The cases were studied using immunohistochemically using alpha-inhibin. In addition, an ovarian tissue microarray containing 170 benign and malignant ovarian neoplasms was also studied immunohistochemically for calretinin expression. The ovarian microarray included 14 SCST, 59 ovarian serous borderline tumors, 17 mucinous borderline tumors, 10 mucinous adenocarcinomas, 32 endometrioid adenocarcinomas, 34 clear cell carcinomas, and 4 germ cell tumors. Results: 99% of SCST examined using full tissue sections exhibited positive cytoplasmic staining for inhibin. On the contrary, only 7% of the histological mimics (P value < 0.0001). 86% of SCST in the tissue microarray were positive for calretinin with nuclear and/or cytoplasmic staining compared to only 7% of the other tumor types (P value < 0.0001). Conclusions: SCST have characteristic clinicopathological and immunohistochemical features and their recognition is crucial for proper diagnosis and treatment. Alpha-inhibin and calretinin are of great help in the diagnosis of sex cord-stromal tumors.

Keywords: calretinin, granulosa cell tumor, inhibin, sex cord-stromal tumors

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Authors: Sundaresan Sivapatham, B. Prabhu

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Cancer results from a multistage, multi-mechanism carcinogenesis process that involves mutagenic, cell death and epigenetic mechanisms, during the three distinguishable but closely allied stages: initiation, promotion, and progression. Chemoprevention is promising in the realm of cancer prevention and it has been shown to reduce the risk of development of carcinoma in highly susceptible individuals such as those with known genetic mutations or high level of risk factors. The present study is aimed at the need of early detection of bladder cancer in order to improve performance in the treatment of this disease. Consumption of certain natural products like DIM is associated with a reduction in cancer incidence in humans. The study showed the protective effects of Diindolylmethane in N-Butyl-N-(4-hydroxybutyl) nitrosamine treated rats. Results of the study had shown the changes in the tumor markers, biomarkers and histopathological alterations in experimental rats when compared to control rats. The protective effects of DIM were shown from the results of cell proliferation, apoptotic markers and histopathological findings when compared with experimental control animals. Hence, our results speculate that the tumor markers, apoptotic markers, histopathological changes and cell proliferation index measured as PCNA serves as an indicator suggestive of protective effects of DIM in BBN induced urinary bladder carcinogenesis.

Keywords: bladder cancer, N-Butyl-N-(4-hydroxybutyl) nitrosamine, diindolylmethane, histopathology

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Authors: Shikha Rani, Neeta Sehgal, Vipin Kumar Verma, Om Prakash

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Introduction: Fish is an important protein source for humans and has great economic value. Fish cultures are affected due to various anthropogenic activities that lead to bacterial and viral infections. Aeromonas hydrophila is a fish pathogenic bacterium that causes several aquaculture outbreaks throughout the world and leads to huge mortalities. In this study, plants of no commercial value were used to investigate their immunostimulatory, antioxidant, anti-inflammatory, anti-bacterial, and disease resistance potential in fish against Aeromonas hydrophila, through fish feed fortification. Methods: The plant was dried at room temperature in the shade, dissolved in methanol, and analysed for biological compounds through GC-MS/MS. DPPH, FRAP, Phenolic, and flavonoids were estimated following standardized protocols. In silico molecular docking was also performed to validate its broad-spectrum activities based on binding affinity with specific proteins. Fish were divided into four groups (n=6; total 30 in a group): Group 1, non-challenged fish (fed on a non-supplemented diet); Group 2, fish challenged with bacteria (fed on a non-supplemented diet); Group 3 and 4, fish challenged with bacteria (A. hydrophila) and fed on plant supplemented feed at 2.5% and 5%. Blood was collected from the fish on 0, 7th, 14th, 21st, and 28th days. Serum was separated for glutamic-oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), alkaline phosphatase assay (ALP), lysozyme activity assay, superoxide dismutase assay (SOD), lipid peroxidation assay (LPO) and molecular parameters (including cytokine levels) were estimated through ELISA. The phagocytic activity of macrophages from the spleen and head kidney, along with quantitative analysis of immune-related genes, were analysed in different tissue samples. The digestive enzymes (Pepsin, Trypsin, and Chymotrypsin) were also measured to evaluate the effect of plant-supplemented feed on freshwater fish. Results and Discussion: GC-MS/MS analysis of a methanolic extract of plant validated the presence of key compounds having antioxidant, anti-inflammatory, anti-bacterial, anti-inflammatory, and immunomodulatory activities along with disease resistance properties. From biochemical investigations like ABTS, DPPH, and FRAP, the amount of total flavonoids, phenols, and promising binding affinities towards different proteins in molecular docking analysis helped us to realize the potential of this plant that can be used for investigation in the supplemented feed of fish. Measurement liver function tests, ALPs, oxidation-antioxidant enzyme concentrations, and immunoglobulin concentrations in the experimental groups (3 and 4) showed significant improvement as compared to the positive control group. The histopathological evaluation of the liver, spleen, and head kidney supports the biochemical findings. The isolated macrophages from the group fed on supplemented feed showed a higher percentage of phagocytosis and a phagocytic index, indicating an enhanced cell-mediated immune response. Significant improvements in digestive enzymes were also observed in fish fed on supplemented feed, even after weekly challenges with bacteria. Hence, the plant-fortified feed can be recommended as a regular feed to enhance fish immunity and disease resistance against the Aeromonas hydrophila infection after confirmation from the field trial.

Keywords: immunostimulation, antipathogen, plant fortified feed, macrophages, GC-MS/MS, in silico molecular docking

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Authors: Mai M. Farid, Mona El-Shabrawy, Sameh R. Hussein, Ahmed Elkhateeb, El-Said S. Abdel-Hameed, Mona M. Marzouk

Abstract:

Asphodelus aestivus Brot. Is a wild plant distributed in Egypt and is considered one of the five Asphodelus spp. from the family Asphodelaceae; it grows in dry grasslands and on rocky or sandy soil. The chemical components of A. aestivus flowers extract were analyzed using different chromatographic and spectral techniques and led to the isolation of two anthraquinones identified as emodin and emodin-O-glucoside. In addition to, five flavonoid compounds;kaempferol,Kaempferol-3-O-glucoside,Apigenin-6-C-glucoside-7-O-glucoside (Saponarine), luteolin 7-O-β-glucopyranoside, Isoorientin-O-malic acid which is a new compound in nature. The LC-ESI-MS/MS analysis of the flower extract of A. aestivus led to the identification of twenty- two compounds characterized by the presence of flavones, flavonols, and flavone C-glycosides. While GC/MS analysis led to the identification of 24 compounds comprising 98.32% of the oil, the major components of the oil were 9, 12, 15-Octadecatrieoic acid methyl ester 28.72%, and 9, 12-Octadecadieroic acid (Z, Z)-methyl ester 19.96%. In vitro cytotoxic activity of the aqueous methanol extract of A. aestivus flowers against HEPG2, HCT-116, MCF-7, and A549 culture was examined and showed moderate inhibition (62.3±1.1)% on HEPG2 cell line followed by (36.8±0.2)% inhibition on HCT-116 and a weak inhibition (5.7± 0.0.2) on MCF-7 cell line followed by (4.5± 0.4) % inhibition on A549 cell line and this is considered the first cytotoxic report of A. aestivus flowers.

Keywords: Anthraquinones, Asphodelus aestivus, Cytotoxic activity, Flavonoids, LC-ESI-MS/MS

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