Search results for: serum biochemical indexes

Authors: Dong-An Wang

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All kinds of microspheres have been extensively employed as carriers for drug, gene and therapeutic cell delivery. Most therapeutic cell delivery microspheres rely on a two-step methodology: fabrication of microspheres and subsequent seeding of cells onto them. In this study, we have developed a novel one-step cell encapsulation technique using a convenient and instant water-in-oil single emulsion approach to form cell-encapsulated gelatin microspheres. This technology is adopted for hyaline cartilage tissue engineering, in which autologous chondrocytes are used as therapeutic cells. Cell viability was maintained throughout and after the microsphere formation (75-100 µm diameters) process that avoids involvement of any covalent bonding reactions or exposure to any further chemicals. Further encapsulation of cell-laden microspheres in alginate gels were performed under 4°C via a prompt process. Upon the formation of alginate constructs, they were immediately relocated into CO2 incubator where the temperature was maintained at 37°C; under this temperature, the cell-laden gelatin microspheres dissolved within hours to yield similarly sized cavities and the chondrocytes were therefore suspended within the cavities inside the alginate gel bulk. Hence, the gelatin cell-laden microspheres served two roles: as cell delivery vehicles which can be removable through temperature curing, and as porogens within an alginate hydrogel construct to provide living space for cell growth and tissue development as well as better permeability for mutual diffusions. These cell-laden microspheres, namely “temperature-cured dissolvable gelatin microsphere based cell carriers” (tDGMCs), were further encapsulated in a chondrocyte-laden alginate scaffold system and analyzed by WST-1, gene expression analyses, biochemical assays, histology and immunochemistry stains. The positive results consistently demonstrated the promise of tDGMC technology in delivering these non-anchorage dependent cells (chondrocytes). It can be further conveniently translated into delivery of other non-anchorage dependent cell species, including stem cells, progenitors or iPS cells, for regeneration of tissues in internal organs, such as engineered hepatogenesis or pancreatic regeneration.

Keywords: biomaterials, tissue engineering, microsphere, hydrogel, porogen, anchorage dependence

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Authors: Pravina Gurjar, Bothiraja Pour, Vijay Kumbhar, Ganesh Dama

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Andrographolide (AG), a bioactive phytoconstituent, has a wider range of pharmacological action. However, due to the intestinal degradation, shows low oral bioavailability. The aim of the present work was to develop Floating In-situ gelling Gastro retentive System (FISGS) for AG in order to enhance its site specific absorption and minimize pH dependent hydrolysis in alkaline environment. Further to increase its therapeutic efficacy for peptic ulcer disease caused by H. pyroli. Gellan based floating in situ gelling system of AG were prepared by using sodium citrate and calcium carbonate. The 32 factorial designs was used to study the effect of gellan and calcium carbonate concentration (independent variables) on dependent variable such as viscosity, floating lag time and drug release. Developed system was evaluated for drug content, floating lag time, viscosity, and drug release studies. Drug content, viscosity, and floating lag time was found to be 81-99%, 67-117 Cps, and 3-5 sec, respectively. The obtained system showed good in vitro floating ability for more than 12 h using 0.1 N HCl as dissolution medium with initial burst release followed by the controlled zero order drug release up to 24 hrs. In vivo testing of FISGS of AG to rats demonstrated significant antiulcer activity that were evaluated by various parameters like pH, volume, total acidity, millimole equivalent of H+ ions/30 min, and protein content of gastric content. The densities of all the formulation batches were found to be near about 0.9 and floating duration above 12 hr. It was observed that with the increase in conc. of gellan there was increase in the viscosity of formulation but all formulations were in optimum range. The drug content of optimized batch was found to be 99.23. In histopathology study of stomach, the villi at the mucosal surface, the intercellular junction, the intestinal lumen were intact; no destruction of the epithelium, and submucosal gland in formulation treated and control group animals as compared to pure drug AG and standard ranitidine. Gellan-based in situ gastro retentive floating system could be advantageous in terms of increased bioavailability of AG to maintain an effective drug conc. in gastric fluid as well as in serum for longer period of time.

Keywords: andrographolide, floating drug delivery, in situ gelling system, gastroretentive system

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Authors: Ali Showail Ali Alasmari

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Introduction: Cataract is the loss of transparency of the lens inside the eye. It is the most common cause of visual loss and blindness worldwide. This study provides a systemic review of the recent findings on the association of vitamin D, and vitamin B12, and their possible role in preventing cataracts in senile (S) and diabetic mellitus (DM) patient groups. Objective: This study was intended to establish and investigate if there is any role between vitamin D and vitamin B12? Secondly, the connection between serum level of vitamin D and vitamin B12 in cataract incidence senile (s) vs. diabetic mellitus (DM) cataract patient groups. Furthermore, to evaluate and analyze cataract occurrence regarding vitamin D and vitamin B12 levels with other risk factors. Finally, to evaluate lens opacities pre and post treatment with vitamin D and vitaminB12 linked to age and visual acuity loss in both senile(S) and diabetic mellitus (DM) cataract patients’ groups. Methods: This study conducted at the ophthalmology clinic at Muhyail General Hospital. Select a prospective case-control to study the effect of vitamin D and Vit B12 on senile(S) cataracts that caused by age and diabetic mellitus (DM)cataract patients; then we compare these two groups. This study prospectively enrolled a total of 50 samples, 25 with senile cataract and 25 with diabetic cataract, from ophthalmology clinic at Muhyail General Hospital. Measuring 25-hydroxy vitamin D and vitamin B12 level concentrations in the assigned samples. Analyses were performed using SAS (statistical analysis software) program. Results: The most important finding in this study was that the senile(s) cataract patients’ group greatly benefited by the combination therapy of vitamin D, and Vitamin B12 reached (28.5±1.50 and 521.1±21.10) respectively; on the contrary, the diabetic cataract patient group hardly shows any significant improvement (21.5 ± 1.00 and 197.2 ± 7.20) respectively. This is because of the Metformin, the first line drug for treating diabetes, has been reported to potentially decrease vitamin B-12 status. This epigenetic modification was correlated with the diabetic mellitus (DM) cataract patients’ group not responding. Vitamin B12 deficiency also leads to an impairment of the conversion of methylmalonyl-CoA to succinyl-CoA, which has been associated with insulin resistance. There was no significant difference between the age, body mass index (BMI), the mean of Vit-D pre-treatments, and the mean values of Hemoglobin A1C of both senile (S) and diabetic mellitus (DM) cataract patient groups. On other hand, there was a highly significant difference between the mean values of glucose levels in both senile (S) and diabetic mellitus (DM) cataract patient groups. Conclusion: Here we conclude that diabetic mellitus (DM) cataract patient group hardly benefited from this combination therapy vitamin D and vitamin B12; on the other hand senile patient group (s) benefited a lot from the therapy.

Keywords: cataract patients, senile, diabetes mellitus, vitamin B12, vitamin D, Muhyail General Hospital, Saudi Arabia

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Authors: Ali Ghorbanipour

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The aim of this study is to investigate the drug resistance pattern and the value of the MIC (minimum inhibitory concentration)method to reduce the impact of infectious diseases and the slow development of resistance. Method: The study was conducted on clinical specimens collected between 2018 to 2021. identification of isolates and antibiotic susceptibility testing were performed using conventional biochemical tests. Antibiotic resistance was determined using kibry-Bauer disk diffusion and MIC by E-test methods comparative with microdilution plate elisa method. Results were interpreted according to CLSI. Results: Out of 249600 different clinical specimens, 18720 different pathogenic bacteria by overall detection ratio 7.7% were detected. Among pathogen bacterial were Gram negative bacteria (70%,n=13000) and Gram positive bacteria(30%,n=5720).Medically relevant gram-negative bacteria include a multitude of species such as E.coli , Klebsiella .spp , Pseudomonas .aeroginosa , Acinetobacter .spp , Enterobacterspp ,and gram positive bacteria Staphylococcus.spp , Enterococcus .spp , Streptococcus .spp was isolated . Conclusion: Our results highlighted that the resistance ratio among Gram Negative bacteria and Gram positive bacteria with different infection is high it suggest constant screening and follow-up programs for the detection of antibiotic resistance and the value of MIC drug susceptibility reporting that provide a new way to the usage of resistant antibiotic in combination with other antibiotics or accurate weight of antibiotics that inhibit or kill bacteria. Evaluation of wrong medication in the expansion of resistance and side effects of over usage antibiotics are goals. Ali ghorbanipour presently working as a supervision at the microbiology department of Massoud medical laboratory. Iran. Earlier, he worked as head department of pulmonary infection in firoozgarhospital, Iran. He received master degree in 2012 from Fergusson College. His research prime objective is a biologic wound dressing .to his credit, he has Published10 articles in various international congresses by presenting posters.

Keywords: antimicrobial profile, MIC & MBC Method, microplate antimicrobial assay, E-test

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Authors: Ishfaq A. Bukhari, Bassem Yousef Sheikh, Abdulrahman Almotrefi, Osama Yousaf, Amer Mahmood

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Olive oil is the primary source of fat in the Mediterranean diet which is associated with a low mortality for cardiovascular disease. Olive oil is rich in monounsaturated fatty acids, and has been reported for variety of beneficial cardiovascular effects including blood pressure lowering, anti-platelet, anti-diabetic and anti-inflammatory effects. Growing number evidences from preclinical and clinical studies have shown that olive oil improves insulin resistance, decrease vessels stiffness and prevent thromboembolism. We evaluated the effects of olive against streptozotocin-induced physiological disorders in the animal models of diabetes and ischemia and reperfusion (I/R)- induced cardiac arrhythmias. Diabetes was induced in male rats with a single intraperitoneal injection of streptozotocin (60 mg/kg), rats were treated for two months with olive oil (1 ml/kg p.o). Control animals received saline. Blood glucose, body weight were monitored every 14 days. At the end of the treatment rats were sacrificed hearts were isolated for mounting on langedorff’s apparatus. The blood glucose and body weight was not significantly different in the control and olive treated animals. The control diabetic animals exhibited 100% incidence of I/R –induced ventricular fibrillation which was reduced to 0% with olive oil, treatment. The duration of ventricular fibrillation reduced from 98.8± 2.3 (control) to 0 seconds in the olive oil treated group. Diltiazem, a calcium channel blocker (1 µm/L) showed similar results and protected the I/R-induced cardiac disorders. The biochemical analysis of the cardiac tissues showed that diabetes and I/R produce marked pathological changes in the cardiomyocytes including decreased glutathione (GSH) and increased oxidative stress (Malondialdehyde; MDA). Pretreatment of animals with olive oil (1 ml/kg p.o) increased GSH and MDA levels. Olive oil also improved the diabetic-induced histopathological changes in the cardiomyocytes. These finding indicates that olive possesses cardiac protective properties. Further studies are under way in our lab to explore the mechanism of the cardio-protective effect of olive oil.

Keywords: diabeties, ischemia-reperfusion, olive oil, rats heart

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Authors: J. Oustric, L. Berti, J. Santini

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Nowadays, the objective of durable agriculture, and in particular organic agriculture, is to reduce the level of fertilizer inputs used in crops. Limiting the quantity of fertilizer inputs would optimize the economical result and minimizing the environmental impact. Nutrient deficiency, particularly of a major nutrient (N, P, and K), can seriously affect fruit production and quality. In citrus crops, rootstock/scion combinations. In citrus crop, scion/rootstock combinations are used frequently to improve tolerance to various abiotic stresses. New rootstocks are needed to respond to these constraints, and the use of new tetraploid rootstocks better adapted to lower nutrient intake could offer a promising way forward. The aim of this work was to determine whether a better tolerance to nutrient deficiency could be observed in a doubled diploid seedling and whether this tolerance could be observed in common clementine scion if used as rootstocks. We selected diploid (CM2x) and doubled diploid (CM4x) Citrumelo 4475 seedlings and common clementine (C) grafted onto Citrumelo 4475 diploid (C/CM2x) and doubled diploid (C/CM4x) rootstocks. Nutrient deficiency effects on the seedlings and scion/rootstock combinations were analyzed by studying anatomical, structural and ultrastructural determinants (chlorosis, stomata, ostiole and cells and their organelles), photosynthetic properties (leaf net photosynthetic rate (Pₙₑₜ), stomatal conductance (gₛ), chlorophyll a fluorescence (Fᵥ/Fₘ)) and oxidative marker (malondialdehyde). Nutrient deficiency affected differently foliar tissues, physiological parameters, and oxidative metabolism in leaves of seedlings depending on their ploidy level and of common clementine scion depending on their rootstocks ploidy level. Both CM4x and C/CM4x presented lower foliar damages (chlorosis, chloroplasts, mitochondria, and plastoglobuli), photosynthesis processes alteration (Pₙₑₜ, gₛ, and Fᵥ/Fₘ), and malondialdehyde accumulation than CM2x and C/CM2x after nutrient deficiency. Doubled diploid Citrumelo 4475 can improve nutrient deficiency tolerance, and its use as a rootstock allows to confer this tolerance to the common clementine scion.

Keywords: nutrient deficiency, oxidative stress, photosynthesis, polyploid rootstocks

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Authors: Varsha Salian, Shama Rao, N. Narendra, B. Mohana Kumar

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Evolving evidence proposes the existence of a highly tumorigenic subpopulation of undifferentiated, self-renewing cancer stem cells, responsible for exhibiting resistance to conventional anti-cancer therapy, recurrence, metastasis and heterogeneous tumor formation. Importantly, the mechanisms exploited by cancer stem cells to resist chemotherapy are very less understood. Oral squamous cell carcinoma (OSCC) is one of the most regularly diagnosed cancer types in India and is associated commonly with alcohol and tobacco use. Therefore, the isolation and in vitro characterization of cancer stem-like cells from patients with OSCC is a critical step to advance the understanding of the chemoresistance processes and for designing therapeutic strategies. With this, the present study aimed to establish and characterize cancer stem-like cells in vitro from OSCC. The primary cultures of cancer stem-like cell lines were established from the tissue biopsies of patients with clinical evidence of an ulceroproliferative lesion and histopathological confirmation of OSCC. The viability of cells assessed by trypan blue exclusion assay showed more than 95% at passage 1 (P1), P2 and P3. Replication rate was performed by plating cells in 12-well plate and counting them at various time points of culture. Cells had a more marked proliferative activity and the average doubling time was less than 20 hrs. After being cultured for 10 to 14 days, cancer stem-like cells gradually aggregated and formed sphere-like bodies. More spheroid bodies were observed when cultured in DMEM/F-12 under low serum conditions. Interestingly, cells with higher proliferative activity had a tendency to form more sphere-like bodies. Expression of specific markers, including membrane proteins or cell enzymes, such as CD24, CD29, CD44, CD133, and aldehyde dehydrogenase 1 (ALDH1) is being explored for further characterization of cancer stem-like cells. To summarize the findings, the establishment of OSCC derived cancer stem-like cells may provide scope for better understanding the cause for recurrence and metastasis in oral epithelial malignancies. Particularly, identification and characterization studies on cancer stem-like cells in Indian population seem to be lacking thus provoking the need for such studies in a population where alcohol consumption and tobacco chewing are major risk habits.

Keywords: cancer stem-like cells, characterization, in vitro, oral squamous cell carcinoma

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Authors: Ofonime Ogba, Augustine Akpan

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Objectives and Goals: The study was aimed at determining pathogenic microbial carriage on the external body parts of Camponotus consobrinus which is also known as the banded sugar ant because of its liking for sugar and sweet food. The level of pathogenic microbial carriage of Camponotus consobrinus in association to the environment in which they have been collected is not known. Methods: The ants were purposively collected from four locations including the kitchens, bedroom of various homes, food shops, and bakeries. The sample collection took place within the hours of 6:30 pm to 11:00 pm. The ants were trapped in transparent plastic containers of which sugar, pineapple peels, sugar cane and soft drinks were used as bait. The ants were removed with a sterile spatula and put in 10mls of peptone water in sterile universal bottles. The containers were vigorously shaken to wash the external surface of the ant. It was left overnight and transported to the Microbiology Laboratory, University of Calabar Teaching Hospital for analysis. The overnight peptone broths were inoculated on Chocolate agar, Blood agar, Cystine Lactose Electrolyte-Deficient agar (CLED) and Sabouraud dextrose agar. Incubation was done aerobically and in a carbon dioxide jar for 24 to 48 hours at 37°C. Isolates were identified based on colonial characteristics, Gram staining, and biochemical tests. Results: Out of the 250 Camponotus consobrinus caught for the study, 90(36.0%) were caught in the kitchen, 75(30.0%) in the bedrooms 40(16.0%) in the bakery while 45(18.0%) were caught in the shops. A total of 82.0% prevalence of different microbial isolates was associated with the ants. The kitchen had the highest number of isolates 75(36.6%) followed by the bedroom 55(26.8%) while the bakery recorded the lowest number of isolates 35(17.1%). The profile of micro-organisms associated with Camponotus consobrinus was Escherichia coli 73(30.0%), Morganella morganii 45(18.0%), Candida species 25(10.0%), Serratia marcescens 10(4.0%) and Citrobacter freundii 10(4.0%). Conclusion: Most of the Camponotus consobrinus examined in the four locations harboured potential pathogens. The presence of ants in homes and shops can facilitate the propagation and spread of pathogenic microorganisms. Therefore, the development of basic preventive measures and the control of ants must be taken seriously.

Keywords: Camponotus consobrinus, potential pathogens, microbial isolates, spread

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Authors: Mohammad Nasb, Muhammad Rehan, Chen Hong

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Background Meniscus defects critically alter knee function and lead to degenerative changes. Regenerative medicine applications including stem cell transplantation have showed a promising efficacy in finding alternatives to overcome traditional treatment limitations. However, stem cell therapy remains limited due to the substantially reduced viability and inhibitory microenvironment. Since tissue growth and repair are under the control of biochemical and mechanical signals, several approaches have recently been investigated (e.g., low intensity pulsed ultrasound [LIPUS]) to promote the regeneration process. This study employed LIPUS to improve growth and osteogenic differentiation of mesenchymal stem cells derived from human embryonic stem cells to improve the regeneration of meniscus tissue. Methodology: The Mesenchymal stromal cells (MSCs) were transplanted into the epicenter of the injured meniscus in rabbits, which were randomized into two main groups: a treatment group (n=32 New Zealand rabbits) including 4 subgroups of 8 rabbits in each subgroup (LIPUS treatment, MSC treatment, LIPUS with MSC and control), and a second group (n=9) to track implanted cells and their progeny using green fluorescence protein (GFP). GFP consists of the MSC and LIPUS-MSC combination subgroups. Rabbits were then subjected to histological, immunohistochemistry, and MRI assessment. Results: The quantity of the newly regenerated tissue in the combination treatment group that had Ultrasound irradiation after mesenchymal stem cells were better at all end points. Likewise, Tissue quality scores were also greater in knees treated with both approaches compared with controls and single treatment at all end points, achieving significance at twelve and twenty-four weeks [p < 0.05], and [p = 0.008] at twelve weeks. Differentiation into type-I and II collagen-expressing cells were higher in the combination group at up to twenty-four weeks. Conclusions: the combination of mesenchymal stem cells and LIPUS showed greater adhering to the sites of meniscus injury, differentiate into cells resembling meniscal fibrochondrocytes, and improve both quality and quantity of meniscal regeneration.

Keywords: stem cells, regenerative medicine, osteoarthritis, knee

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Authors: Temesgen Sidamo, Prakruti S. Rao, Eleni Akllilu, Workineh Shibeshi, Yumi Park, Yong-Soon Cho, Jae-Gook Shin, Scott K. Heysell, Stellah G. Mpagama, Ephrem Engidawork

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The fluoroquinolones (FQs) are used off-label for the treatment of multidrug-resistant tuberculosis (MDR-TB), and for evaluation in shortening the duration of drug-susceptible TB in recently prioritized regimens. Within the class, levofloxacin (LFX) and moxifloxacin (MXF) play a substantial role in ensuring success in treatment outcomes. However, sub-therapeutic plasma concentrations of either LFX or MXF may drive unfavorable treatment outcomes. To the best of our knowledge, the pharmacokinetics of LFX and MXF in Ethiopian patients with MDR-TB have not yet been investigated. Therefore, the aim of this study was to develop a population pharmacokinetic (PopPK) model of levofloxacin (LFX) and moxifloxacin (MXF) and assess the percent probability of target attainment (PTA) as defined by the ratio of the area under the plasma concentration-time curve over 24-h (AUC0-24) and the in vitro minimum inhibitory concentration (MIC) (AUC0-24/MIC) in Ethiopian MDR-TB patients. Steady-state plasma was collected from 39 MDR-TB patients enrolled in the programmatic treatment course and the drug concentrations were determined using optimized liquid chromatography-tandem mass spectrometry. In addition, the in vitro MIC of the patients' pretreatment clinical isolates was determined. PopPK and simulations were run at various doses, and PK parameters were estimated. The effect of covariates on the PK parameters and the PTA for maximum mycobacterial kill and resistance prevention was also investigated. LFX and MXF both fit in a one-compartment model with adjustments. The apparent volume of distribution (V) and clearance (CL) of LFX were influenced by serum creatinine (Scr), whereas the absorption constant (Ka) and V of MXF were influenced by Scr and BMI, respectively. The PTA for LFX maximal mycobacterial kill at the critical MIC of 0.5 mg/L was 29%, 62%, and 95% with the simulated 750 mg, 1000 mg, and 1500 mg doses, respectively, whereas the PTA for resistance prevention at 1500 mg was only 4.8%, with none of the lower doses achieving this target. At the critical MIC of 0.25 mg/L, there was no difference in the PTA (94.4%) for maximum bacterial kill among the simulated doses of MXF (600 mg, 800 mg, and 1000 mg), but the PTA for resistance prevention improved proportionately with dose. Standard LFX and MXF doses may not provide adequate drug exposure. LFX PopPK is more predictable for maximum mycobacterial kill, whereas MXF's resistance prevention target increases with dose. Scr and BMI are likely to be important covariates in dose optimization or therapeutic drug monitoring (TDM) studies in Ethiopian patients.

Keywords: population PK, PTA, moxifloxacin, levofloxacin, MDR-TB patients, ethiopia

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Authors: Hamada Ahmed, Mervat A. Abdel-Latif, Alaa. A. Ghoraba, Samah A. Ganna

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An experiment was conducted to determine the effect of microbial phytase (Quantum Blue®) supplementation on layer chickens fed diets with required or low phosphorous level in corn-soybean based diets. One hundred and sixteen 23-week-old Lohman brown laying hens were used in 8-week feeding trial. Hens were randomly allotted into four treatments where the group (1) (control group) was fed basal diet without phytase, group (2) fed basal diet supplemented with phytase, group (3) fed diet supplemented with phytase as a replacement of 25% of monocalcium phosphate and group (4) fed diet supplemented with phytase as a replacement of 50% of monocalcium phosphate. Records on daily egg production, egg mass, egg weight and body weight of hens at the end of experimental period were recorded. Results revealed no significant (p ≥ 0.05) differences were observed among the other dietary treatments in BW, egg production, egg mass, feed intake or feed conversion when these parameters were evaluated over the duration of the experiment while egg weight showed significant (p < 0.05) increase in all phytase supplemented groups. There was no significant (p ≥ 0.05) differences in egg quality including egg length, egg width, egg shape index, yolk height, yolk width, yolk index, yolk weight and yolk albumin ratio while egg albumin was significantly increased (p < 0.05) in group (2) and group (3). Egg shell weight increased significantly (p < 0.05) in all phytase supplemented groups when compared with the control group also shell thickness increased significantly (p < 0.05) in both group (2 &3). No significant (P ≥ 0.05) difference was observed in serum Ca, P level while alkaline phosphatase was significantly (P ˂ 0.05) increased in group (3). Egg shell analysis showed increase in egg shell ash% in all phytase supplemented groups when compared with the control group, egg shell calcium % was higher in group (3) and group (4) than the control group while group (2) showed lower egg shell calcium% than the other experimental groups, egg shell phosphorous% was higher in all phytase supplemented groups than the control group. Phosphorous digestability was significantly (P ˂ 0.05) increased in all phytase supplemented groups than the control group and the highest p digestability was in group (4). Calcium digestability showed significant (P ˂ 0.05) increase in all phytase supplemented groups when compared with the control group and the highest digetability was in group (4).

Keywords: layers, microbial phytase, Ca and P availability, egg production, egg characteristics

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Authors: Himani Sharma, Amit Agrawal

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Stability of the Cassie and Wenzel wetting states depends on intrinsic contact angle and geometric features on a surface that was exploited in capturing biofluids in microwells. However, the mechanism of imbibition of biofluids in the microwells is not well implied in terms of wettability of a substrate. In this work, we experimentally demonstrated filling dynamics in hydrophilic and hydrophobic microwells by protein solutions. Towards this, we utilized lotus leaf as a mold to fabricate microwells on a Polydimethylsiloxane (PDMS) surface. Lotus leaf containing micrometer-sized blunt-conical shaped pillars with a height of 8-15 µm and diameter of 3-8 µm were transferred on to PDMS. Furthermore, PDMS surface was treated with oxygen plasma to render the hydrophilic nature. A 10µL droplets containing fluorescein isothiocyanate (FITC) - labelled bovine serum albumin (BSA) were rested on both hydrophobic (θa = 108o, where θa is the apparent contact angle) and hydrophilic (θa = 60o) PDMS surfaces. A time-dependent fluorescence microscopy was conducted on these modified PDMS surfaces by recording the fluorescent intensity over a 5 minute period. It was observed that, initially (at t=1 min) FITC-BSA was accumulated on the periphery of both hydrophilic and hydrophobic microwells due to incomplete penetration of liquid-gas meniscus. This deposition of FITC-BSA on periphery of microwell was not changed with time for hydrophobic surfaces, whereas, a complete filling was occurred in hydrophilic microwells (at t=5 mins). This attributes to a gradual movement of three-phase contact line along the vertical surface of the hydrophilic microwells as compared to stable pinning in the hydrophobic microwells as confirmed by Surface Evolver simulations. In addition, if the cavities are presented on hydrophobic surfaces, air bubbles will be trapped inside the cavities once the aqueous solution is placed over these surfaces, resulting in the Cassie-Baxter wetting state. This condition hinders trapping of proteins inside the microwells. Thus, it is necessary to impart hydrophilicity to the microwell surfaces so as to induce the Wenzel state, such that, an entire solution will be fully in contact with the walls of microwells. Imbibition of microwells by protein solutions was analyzed in terms fluorescent intensity versus time. The present work underlines the importance of geometry of microwells and surface wettability of substrate in wetting and effective capturing of solid sub-phases in biofluids.

Keywords: BSA, microwells, surface evolver, wettability

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Authors: Mustafa M. Donma, Ayşen Haksayar, Bahadır Batar, Buse Tepe, Birol Topçu, Orkide Donma

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Childhood obesity is an ever-increasing health problem. The Association of obesity with severe chronic diseases such as diabetes and cardiovascular diseases makes the problem life-threatening. Aside from psychological, societal and metabolic factors, genetic polymorphisms have gained importance concerning etiology in recent years. The aim of this study was to evaluate the relationship between rs10455872 gene polymorphism in the Lipoprotein (a) locus and the development of childhood obesity. This was a prospective study carried out according to the Helsinki Declarations. The study protocol was approved by the Institutional Ethics Committee. This study was supported by Tekirdag Namik Kemal University Rectorate, Scientific Research Projects Coordination Unit. Project No: NKUBAP.02.TU.20.278. A total of 180 children (103 obese (OB) and 77 healthy), aged 6-18 years, without any acute or chronic disease, participated in the study. Two different groups were created: OB and healthy control. Each group was divided into two further groups depending on the nature of the polymorphism. Anthropometric measurements were taken during the detailed physical examination. Laboratory tests and TANITA measurements were performed. For the statistical evaluations, SPSS version 28.0 was used. A P-value smaller than 0.05 was the statistical significance degree. The distribution of lipoprotein (a) rs10455872 gene polymorphism did not differ between OB and healthy children. Children with AG genotype in both OB and control groups had lower body mass index (BMI), diagnostic obesity notation model assessment index (DONMA II), body fat ratio (BFR), C-reactive protein (CRP), and metabolic syndrome index (MetS index) values compared to children with normal AA genotype. In the OB group, serum iron, vitamin B12, hemoglobin, MCV, and MCH values were found to be higher in the AG genotype group than those of children with the normal AA genotype. A significant correlation was found between the MetS index and BFR among OB children with normal homozygous genotype. MetS index increased as BFR increased in this group. However, such a correlation was not observed in the OB group with heterozygous AG genotype. To the best of our knowledge, the association of lipoprotein (a) rs10455872 gene polymorphism with the etiology of childhood obesity has not been studied yet. Therefore, this study was the first report suggesting polymorphism with AG genotype as a good risk factor for obesity.

Keywords: child, gene polymorphism, lipoprotein (a), obesity, rs10455872

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Authors: Vincenzo Piemonte

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The liver is a complex, large-scale biochemical reactor which plays a unique role in the human physiology. When liver ceases to perform its physiological activity, a functional replacement is required. Actually, liver transplantation is the only clinically effective method of treating severe liver disease. Anyway, the aforementioned therapeutic approach is hampered by the disparity between organ availability and the number of patients on the waiting list. In order to overcome this critical issue, research activities focused on liver support device systems (LSDs) designed to bridging patients to transplantation or to keep them alive until the recovery of native liver function. In recirculating albumin dialysis devices, such as MARS (Molecular Adsorbed Recirculating System), adsorption is one of the fundamental steps in albumin-dialysate regeneration. Among the albumin-bound toxins that must be removed from blood during liver-failure therapy, bilirubin and tryptophan can be considered as representative of two different toxin classes. The first one, not water soluble at physiological blood pH and strongly bounded to albumin, the second one, loosely albumin bound and partially water soluble at pH 7.4. Fixed bed units are normally used for this task, and the design of such units requires information both on toxin adsorption equilibrium and kinetics. The most common adsorptive media used in LSDs are activated carbon, non-ionic polymeric resins and anionic resins. In this paper, bilirubin adsorption isotherms on different adsorptive media, such as polymeric resin, albumin-coated resin, anionic resin, activated carbon and alginate beads with entrapped albumin are presented. By comparing all the results, it can be stated that the adsorption capacity for bilirubin of the five different media increases in the following order: Alginate beads < Polymeric resin < Albumin-coated resin < Activated carbon < Anionic resin. The main focus of this paper is to provide useful guidelines for the optimization of liver support devices which implement adsorption columns to remove albumin-bound toxins from albumin dialysate solutions.

Keywords: adsorptive media, adsorption equilibrium, artificial liver devices, bilirubin, mathematical modelling

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Authors: Dany I. Doughan, Nizam S. Najd

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Different governments in today’s global pandemic are approaching the challenging and complex issue of mitigating the spread of the SARS-CoV-2 virus differently while simultaneously considering their national economic and operational bottom lines. One of the most notable successes has been Taiwan's multifaceted virus containment approach, which resulted in a substantially lower incidence rate compared to Sweden’s chief mitigation tactic of herd immunity. From a classic Swiss Cheese Model perspective, integrating more fail-safe layers of defense against the virus in Taiwan’s approach compared to Sweden’s meant that in Taiwan, the government did not have to resort to extreme measures like the national lockdown Sweden is currently contemplating. From an optimized virus spread mitigation solution development standpoint using the Solutions Principle, the Taiwanese and Swedish solutions were desirable economically by businesses that remained open and non-economically or socially by individuals who enjoyed fewer disruptions from what they considered normal before the pandemic. Out of the two, the Taiwanese approach was more feasible long-term from a workforce management and quality control perspective for healthcare facilities and their professionals who were able to provide better, longer, more attentive care to the fewer new positive COVID-19 cases. Furthermore, the Taiwanese approach was more applicable as an overall model to emulate thanks in part to its short-term and long-term multilayered approach, which allows for the kind of flexibility needed by other governments to fully or partially adapt or adopt said, model. The Swedish approach, on the other hand, ignored the biochemical nature of the virus and relied heavily on short-term personal behavioral adjustments and conduct modifications, which are not as reliable as establishing required societal norms and awareness programs. The available international data on COVID-19 cases and the published governmental approaches to control the spread of the coronavirus support a better fit into the Solutions Principle of Taiwan’s Swiss Cheese Model success story compared to Sweden’s.

Keywords: coronavirus containment and mitigation, solutions principle, Swiss Cheese Model, viral mutation

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Authors: Noura El-Ahmady El-Naggar

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L-asparaginase is an important enzyme as therapeutic agents used in combination therapy with other drugs in the treatment of acute lymphoblastic leukemia in children. L-asparaginase producing actinomycete strain, NEAE-K, was isolated from soil sample and identified on the basis of morphological, cultural, physiological and biochemical properties, together with 16S rDNA sequence as new subsp. Streptomyces rochei subsp. chromatogenes NEAE-K and sequencing product (1532 bp) was deposited in the GenBank database under accession number KJ200343. The study was conducted to screen parameters affecting the production of L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K on solid state fermentation using Plackett–Burman experimental design. Sixteen different independent variables including incubation time, moisture content, inoculum size, temperature, pH, soybean meal+ wheat bran, dextrose, fructose, L-asparagine, yeast extract, KNO3, K2HPO4, MgSO4.7H2O, NaCl, FeSO4. 7H2O, CaCl2, and three dummy variables were screened in Plackett–Burman experimental design of 20 trials. The most significant independent variables affecting enzyme production (dextrose, L-asparagine and K2HPO4) were further optimized by the central composite design. As a result, a medium of the following formula is the optimum for producing an extracellular L-asparaginase by Streptomyces rochei subsp. chromatogenes NEAE-K from solid state fermentation: g/L (soybean meal+ wheat bran 15, dextrose 3, fructose 4, L-asparagine 8, yeast extract 2, KNO3 1, K2HPO4 2, MgSO4.7H2O 0.5, NaCl 0.1, FeSO4. 7H2O 0.02, CaCl2 0.01), incubation time 7 days, moisture content 50%, inoculum size 3 mL, temperature 30°C, pH 8.5.

Keywords: streptomyces rochei subsp. chromatogenes neae-k, 16s rrna, identification, solid state fermentation, l-asparaginase production, plackett-burman design, central composite design

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Authors: H. M. El-Rafie, A. H. Abou Zeid, R. S. Mohammed, A. A. Sleem

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Acrocarpus is a genus of flowering plants in the legume family Fabaceae which considered as a large and economically important family. This study aimed to investigate the phytoconstituents of the petroleum ether extract (PEE) of Acrocarpus fraxinofolius bark by Gas chromatography coupled with mass spectrometry (GC/MS) analysis of its fractions (fatty acid and unsaponifiable matter). Concerning this, identification of 52 compounds constituting 97.03 % of the total composition of the unsaponifiable matter fraction. Cycloeucalenol was found to be the major compound representing 32.52% followed by 4a, 14a-dimethyl-A8~24(28)-ergostadien (26.50%) and ß-sitosterol(13.74%), furthermore Gas liquid chromatography (GLC) analysis of the sterol fraction revealed the identification of cholesterol (7.22 %), campesterol (13.30 %), stigmasterol (10.00 %) and β - sitosterol (69.48 %). Meanwhile, the identification of 33 fatty acids representing 90.71% of the total fatty acid constituents. Methyl-9,12-octadecadienoate (40.39%) followed by methyl hexadecanoate (23.64%) were found to be the major compounds. On the other hand, column chromatography and Thin layer chromatography (TLC) fractionation of PEE separate the triterpenoid: 21β-hydroxylup-20(29)-en-3-one and β- amyrin which were structurally identified by spectroscopic analysis (NMR, MS and IR). PEE has been biologically evaluated for 1: management of diabetes in alloxan induced diabetic rats 2: cytotoxic activity against four human tumor cell lines (Cervix carcinoma cell line[HELA], Breast carcinoma cell line [MCF7], Liver carcinoma cell line[HEPG2] and Colon carcinoma cell line[HCT-116] 3: hepatoprotective activity against CCl4-induced hepatotoxicity in rats and the activity was studied by assaying the serum marker enzymes like AST, ALT, and ALP. Concerning this, the anti-diabetic activity exhibited by 100mg of PEE extract was 74.38% relative to metformin (100% potency). It also showed a significant anti-proliferative activity against MCF-7 (IC50= 2.35µg), Hela(IC50=3.85µg) and HEPG-2 (IC50= 9.54µg) compared with Doxorubicin as reference drug. The hepatoprotective activity was evidenced by significant decrease in liver function enzymes, i.e. AST, ALT and ALP by (29.18%, 28.26%, and 34.11%, respectively using silymarin as the reference drug, compared to their concentration levels in an untreated group with liver damage induced by CCl₄. This study was performed for the first time on the bark of this species.

Keywords: Acrocarpus fraxinofolius, antidiabetic, cytotoxic, hepatoprotective

Procedia PDF Downloads 185

Authors: Matthew Ocheleka Itodo, Ogo Agbo Ogo, Agnes Ogbene Abutu, Bawa Inalegwu

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Hyperlipidemia is characterised by elevated serum total cholesterol and low density and very low-density lipoprotein cholesterol and decreased high-density lipoprotein are the risk factor for coronary heart diseases. There are claims by traditional medicine practitioners in Nigeria that Moringa oleifera plants are used for the treatment of cardiovascular diseases, but it appears there is no scientific research and, publication or documented work to verify these claims. This study aimed to determine the effect of methanol root extracts of Moringa oleifera on Lipid profile, Atherogenic indices and 3 hydroxyl 3 methylglutaryl Coenzyme A reductase activity of poloxamer 407-induced hyperlipidemic rats. The animals were grouped into 8; Group 1: Normal control, Group 2: Hyperlipidemic control. Groups 2 to 8 were induced with Poloxamer 407 1000 mg/Kg body weight. However, group 3 were treated with standard drugs (atorvastatin). Group 4 was treated with crude extract, and groups 5 to 8 were treated with purified fractions from column chromatography. The preliminary antihyperlipidemic study showed Methanol root extract at 200 mg/kg body weight significantly (p≤0.05) decreased total cholesterol, low-density lipoprotein, triacylglyceride, 3 hydroxyls 3 methylglutaryl Coenzyme A reductase, and increase high-density lipoprotein of hyperlipidemic treated groups. Screening the extracts for the most potent anti-hyperlipidemic activity reveals that fraction 1 of Total Cholesterol and Fraction 3 of Triacylglyceride have the highest percentage reduction of 56% and 51%, respectively. The atherogenic risk factor of all induced treated rats shows a significant (p<0.05) decrease in levels of Castelli’s risk index II, atherogenic index of plasma and a significant (p<0.05) higher level of Castelli’s risk index I ratio. The study shows that the methanol extract of root possesses antihyperlipidemic effects and may explain why it has been found to be useful in the management of cardiovascular diseases by traditional medicine practitioners.

Keywords: hyperlipidemia, moringa oleifera, poloxamer 407, lipid profile

Procedia PDF Downloads 64

Authors: Zhen Cao, Yu Zhu, Junxue Fu

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Immunoassay, i.e., antigen-antibody reaction, is crucial for disease diagnostics. To achieve the adequate signal of the antigen protein detection, a large amount of sample and long incubation time is needed. However, the amount of protein is usually small at the early stage, which makes it difficult to detect. Unlike cells and DNAs, no valid chemical method exists for protein amplification. Thus, an alternative way to improve the signal is through particle manipulation techniques to concentrate proteins, among which dielectrophoresis (DEP) is an effective one. DEP is a technique that concentrates particles to the designated region through a force created by the gradient in a non-uniform electric field. Since DEP force is proportional to the cube of particle size and square of electric field gradient, it is relatively easy to capture larger particles such as cells. For smaller ones like proteins, a super high gradient is then required. In this work, three-dimensional Ag/SiO2 nanorods arrays, fabricated by an easy physical vapor deposition technique called as oblique angle deposition, have been integrated with a DEP device and created the field gradient as high as of 2.6×10²⁴ V²/m³. The nanorods based DEP device is able to enrich bovine serum albumin (BSA) protein by 1800-fold and the rate has reached 180-fold/s when only applying 5 V electric potential. Based on the above nanorods integrated DEP platform, an immunoassay of mouse immunoglobulin G (IgG) proteins has been performed. Briefly, specific antibodies are immobilized onto nanorods, then IgG proteins are concentrated and captured, and finally, the signal from fluorescence-labelled antibodies are detected. The limit of detection (LoD) is measured as 275.3 fg/mL (~1.8 fM), which is a 20,000-fold enhancement compared with identical assays performed on blank glass plates. Further, prostate-specific antigen (PSA), which is a cancer biomarker for diagnosis of prostate cancer after radical prostatectomy, is also quantified with a LoD as low as 2.6 pg/mL. The time to signal saturation has been significantly reduced to one minute. In summary, together with an easy nanorod fabrication and integration method, this nanorods based DEP platform has demonstrated highly sensitive immunoassay performance and thus poses great potentials in applications for early point-of-care diagnostics.

Keywords: dielectrophoresis, immunoassay, oblique angle deposition, protein concentration

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Authors: Natalia Moreno-Castellanos, Amaia Rodriguez, Gema Frühbeck

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Introduction: In adipocytes, the cytoskeleton undergoes important expression and distribution in adipocytes rearrangements during adipogenesis and in obesity. Indeed, a role for these proteins in the regulation of adipocyte differentiation and response to insulin has been demonstrated. Recently, septins have been considered as new components of the cytoskeletal network that interact with other cytoskeletal elements (actin and tubulin) profoundly modifying their dynamics. However, these proteins have not been characterized as yet in adipose tissue. In this work, were examined the cellular, molecular and functional features of a member of this family, septin 11 (SEPT11), in adipocytes and evaluated the impact of obesity on the expression of this protein in human adipose tissue. Methods: Adipose gene and protein expression levels of SEPT11 were analysed in human samples. SEPT11 distribution was evaluated by immunocytochemistry, electronic microscopy, and subcellular fractionation techniques. GST-pull down, immunoprecipitation and a Yeast-Two Hybrid (Y2H) screening were used to identify the SEPT11 interactome. Gene silencing was employed to assess the role of SEPT11 in the regulation of insulin signaling and lipid metabolism in adipocytes. Results: SEPT11 is expressed in human adipocytes, and its levels increased in both omental and subcutaneous adipose tissue in obesity, with SEPT11 mRNA content positively correlating with parameters of insulin resistance in subcutaneous fat. In non-stimulated adipocytes, SEPT11 immunoreactivity showed a ring-like distribution at the cell surface and associated to caveolae. Biochemical analyses showed that SEPT11 interacted with the main component of caveolae, caveolin-1 (CAV1) as well as with the fatty acid-binding protein, FABP5. Notably, the three proteins redistributed and co-localized at the surface of lipid droplets upon exposure of adipocytes to oleate. In this line, SEPT11 silencing in 3T3-L1 adipocytes impaired insulin signaling and decreased insulin-induced lipogenesis. Conclusions: Those findings demonstrate that SEPT11 is a novel component of the adipocyte cytoskeleton that plays an important role in the regulation of lipid traffic, metabolism and can thus represent a potential biomarker of insulin resistance in obesity in adipocytes through its interaction with both CAV1 and FABP5.

Keywords: caveolae, lipid metabolism, obesity, septins

Procedia PDF Downloads 191

Authors: S. Koppikar, P. Kulkarni, D. Ingale , N. Wagh, S. Deshpande, A. Mahajan, A. Harsulkar

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Crucial role of reactive oxygen species (ROS) in the progression Osteoarthritis (OA) pathogenesis has been endorsed several times though its exact mechanism remains unclear. Oxidative stress is known to instigate classical stress factors such as cytokines, chemokines and ROS, which hampers cartilage remodelling process and ultimately results in worsening the disease. Synovial fluid (SF) is a biological communicator between cartilage and synovium that accumulates redox and biochemical signalling mediators. The present work attempts to measure several oxidative stress markers in the synovial fluid obtained from knee OA patients with varying degree of disease severity. Thirty OA and five Meniscal-tear (MT) patients were graded using Kellgren-Lawrence scale and assessed for Nitric oxide (NO), Nitrate-Nitrite (NN), 2,2-diphenyl-1-picrylhydrazyl (DPPH), Ferric Reducing Antioxidant Potential (FRAP), Catalase (CAT), Superoxide dismutase (SOD) and Malondialdehyde (MDA) levels for comparison. Out of various oxidative markers studied, NO and SOD showed significant difference between moderate and severe OA (p= 0.007 and p= 0.08, respectively), whereas CAT demonstrated significant difference between MT and mild group (p= 0.07). Interestingly, NN revealed statistically positive correlation with OA severity (p= 0.001 and p= 0.003). MDA, a lipid peroxidation by-product was estimated maximum in early OA when compared to MT (p= 0.06). However, FRAP did not show any correlation with OA severity or MT control. NO is an essential bio-regulatory molecule essential for several physiological processes, and inflammatory conditions. However, due to its short life, exact estimation of NO becomes difficult. NO and its measurable stable products are still it is considered as one of the important biomarker of oxidative damage. Levels of NO and nitrite-nitrate in SF of patients with OA indicated its involvement in the disease progression. When SF groups were compared, a significant correlation among moderate, mild and MT groups was established. To summarize, present data illustrated higher levels of NO, SOD, CAT, DPPH and MDA in early OA in comparison with MT, as a control group. NN had emerged as a prognostic bio marker in knee OA patients, which may act as futuristic targets in OA treatment.

Keywords: antioxidant, knee osteoarthritis, oxidative stress, synovial fluid

Procedia PDF Downloads 467

Authors: T. Rakhi, Thrivikram Shenoy

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Esophageal atresia is a disorder of maldevelopment of esophagus with or without a connection to the trachea. Radiological reviews are needed in consultation with the pediatric surgeon and neonatologist and we report a rare case of esophageal atresia associated with atrial septal defect-patent ductus arteriosus complex. A 2-day old female baby born at term, weighing 3.010kg, admitted to the Neonatal Intensive Care Unit with respiratory distress and excessive oral secretions. On examination, continuous murmur and cyanosis were seen. Esophageal atresia was suspected, after a failed attempt to pass a nasogastric tube. Chest radiograph showed coiling of the nasogastric tube and absent gas shadow in the abdomen. Echocardiography confirmed Patent Ductus Arteriosus with Atrial Septal Defect not in failure and was diagnosed with esophageal atresia with suspected fistula posted for surgical repair. After preliminary management with oxygenation, suctioning in prone position and antibiotics, investigations revealed Hb 17gms serum biochemistry, coagulation profile and C-Reactive Protein Test normal. The baby was premedicated with 5mcg of fentanyl and 100 mcg of midazolam and a rapid awake laryngoscopy was done to rule out difficult airway followed by induction with o2 air, sevo and atracurium 2 mg. Placement of a 3.5 tube was uneventful at first attempt and after confirming bilateral air entry positioned in the lateral position for Right thoracotomy. A pulse oximeter, Echocardiogram, Non-invasive Blood Pressure, temperature and a precordial stethoscope in left axilla were essential monitors. During thoracotomy, both the ends of the esophagus and the fistula could not be located after thorough search suggesting an on table finding of type A esophageal atresia. The baby was repositioned for gastrostomy, and cervical esophagostomy ventilated overnight and extubated uneventful. Absent gas shadow was overlooked and the purpose of this presentation is to create an awareness between the neonatologist, pediatric surgeons and anesthesiologist regarding variation of typing of Tracheoesophageal fistula pre and intraoperatively. A need for imaging modalities warranted for a definitive diagnosis in the presence of a gasless stomach.

Keywords: anesthetic, atrial septal defects, esophageal atresia, patent ductus arteriosus, perioperative, chest x-ray

Procedia PDF Downloads 167

Authors: Adriana Garcia-Gurrola, Carlos Adrian Peña Natividad, Ana Laura Martinez Martinez, Alberto Abraham Escobar Puentes, Estefania Ochoa Ruiz, Aracely Serrano Medina, Abraham Wall Medrano, Simon Yobanny Reyes Lopez

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Type 2 diabetes mellitus (T2DM) is a metabolic disease characterized by elevated blood glucose levels. Quercetin is a natural flavonoid with a hypoglycemic effect, but reported data are inconsistent due mainly to the structural instability and low solubility of quercetin. Nanoencapsulation is a distinct strategy to overcome the intrinsic limitations of quercetin. Therefore, this work aims to develop a quercetin nano-formulation based on biopolymeric starch nanoparticles to enhance the release and hypoglycemic effect of quercetin in T2DM induced mice model. Starch-quercetin nanoparticles were synthesized using high-intensity ultrasonication, and structural and colloidal properties were determined by FTIR and DLS. For in vivo studies, CD1 male mice (n=25) were divided into five groups (n=5). T2DM was induced using a high-fat and high-sugar diet for 32 weeks and streptozotocin injection. Group 1 consisted of healthy mice fed with a normal diet and water ad libitum; Group 2 were diabetic mice treated with saline solution; Group 3 were diabetic mice treated with glibenclamide; Group 4 were diabetic mice treated with empty nanoparticles; and Group 5 was diabetic mice treated with quercetin nanoparticles. Quercetin nanoparticles had a hydrodynamic size of 232 ± 88.45 nm, a PDI of 0.310 ± 0.04 and a zeta potential of -4 ± 0.85 mV. The encapsulation efficiency of nanoparticles was 58 ± 3.33 %. No significant differences (p = > 0.05) were observed in biochemical parameters (lipids, insulin, and peptide C). Groups 3 and 5 showed a similar hypoglycemic effect, but quercetin nanoparticles showed a longer-lasting effect. Histopathological studies reveal that T2DM mice groups showed degenerated and fatty liver tissue; however, a treated group with quercetin nanoparticles showed liver tissue like that of the healthy mice group. These results demonstrate that quercetin nano-formulations based on starch nanoparticles are effective alternatives with hypoglycemic effects.

Keywords: quercetin, diabetes mellitus tipo 2, in vivo study, nanoparticles

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Authors: Muhammad Aown Sammar Raza

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Environmental stresses are one of the major reasons for poor crop yield across the globe. Among the various environmental stresses, drought stress is the most damaging one, especially in arid and semi-arid regions. Wheat is the major staple food of many countries of the world, which is badly affected by drought stress. In order to fulfill the dietary needs of increasing population with depleting water resources there is a need to adopt technologies which result in sufficient crop yield with less water consumption. One of them is partial root zone drying. Keeping in view these conditions, a wire house experiment was conducted at agronomic research area of University College of Agriculture and Environmental Sciences, The Islamia University Bahawalpur during 2015, to screen out the different wheat varieties for partial root zone drying (PRD). Five approved local wheat varieties (V1= Galaxy-2013, V2= Punjab-2011, V3 = Faisalabad-2008, V4 = Lasani-2008 and V5 = V.8200) and two irrigation levels (I1= control irrigation and I2 = PRD irrigation) with completely randomized design having four replications were used in the experiment. Among the varieties, Galaxy-2013 performed the best and attained maximum plant height, leaf area, stomatal conductance, photosynthesis, total sugars, proline contents and antioxidant enzymes activities and minimum values of growth and physiological parameters were recorded in variety V.8200. For irrigation levels, higher values of growth, physiological and water related parameters were recorded in control treatment (I1) except leaf water potential, osmotic potential, total sugars and proline contents. However, enzyme activities were higher under PRD treatment for all varieties. It was concluded that Galaxy-2013 is the most compatible and V.8200 is the most susceptible variety for PRD, respectively and more quality traits and enzymatic activities were recorded under PRD irrigation as compared to control treatment.

Keywords: antioxidant enzymes activities, osmolytes concentration, partial root zone drying, photosynthetic rate, water relations, wheat

Procedia PDF Downloads 229

Authors: Aleya Ferdausi, Meriel Jones, Anthony Halls

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The Amaryllidaceae genus Narcissus contains secondary metabolites, which are important sources of bioactive compounds such as pharmaceuticals indicating that their biological activity extends from the native plant to humans. Transcriptome analysis (RNA-seq) is an effective platform for the identification and functional characterization of candidate genes as well as to identify genes encoding uncharacterized enzymes. The biotechnological production of secondary metabolites in plant cell or organ cultures has become a tempting alternative to the extraction of whole plant material. The biochemical pathways for the production of secondary metabolites require primary metabolites to undergo a series of modifications catalyzed by enzymes such as cytochrome P450s, methyltransferases, glycosyltransferases, and acyltransferases. Differential gene expression analysis of Narcissus was obtained from two conditions, i.e. field and in vitro callus. Callus was obtained from modified MS (Murashige and Skoog) media supplemented with growth regulators and twin-scale explants from Narcissus cv. Carlton bulb. A total of 2153 differentially expressed transcripts were detected in Narcissus bulb and in vitro callus, and 78.95% of those were annotated. It showed the expression of genes involved in the biosynthesis of alkaloids were present in both conditions i.e. cytochrome P450s, O-methyltransferase (OMTs), NADP/NADPH dehydrogenases or reductases, SAM-synthetases or decarboxylases, 3-ketoacyl-CoA, acyl-CoA, cinnamoyl-CoA, cinnamate 4-hydroxylase, alcohol dehydrogenase, caffeic acid, N-methyltransferase, and NADPH-cytochrome P450s. However, cytochrome P450s and OMTs involved in the later stage of Amaryllidaceae alkaloids biosynthesis were mainly up-regulated in field samples. Whereas, the enzymes involved in initial biosynthetic pathways i.e. fructose biphosphate adolase, aminotransferases, dehydrogenases, hydroxyl methyl glutarate and glutamate synthase leading to the biosynthesis of precursors; tyrosine, phenylalanine and tryptophan for secondary metabolites were up-regulated in callus. The knowledge of probable genes involved in secondary metabolism and their regulation in different tissues will provide insight into the Narcissus plant biology related to alkaloid production.

Keywords: narcissus, callus, transcriptomics, secondary metabolites

Procedia PDF Downloads 134

Authors: Abdulqader Alkhouzaam, Hazim Qiblawey, Majeda Khraisheh

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Membrane treatment for desalination and wastewater treatment is one of the promising solutions to affordable clean water. It is a developing technology throughout the world and considered as the most effective and economical method available. However, the limitations of membranes’ mechanical and chemical properties restrict their industrial applications. Hence, developing novel membranes was the focus of most studies in the water treatment and desalination sector to find new materials that can improve the separation efficiency while reducing membrane fouling, which is the most important challenge in this field. Graphene oxide (GO) is one of the materials that have been recently investigated in the membrane water treatment sector. In this work, ultrafiltration polysulfone (PSF) membranes with high antifouling properties were synthesized by incorporating different loadings of GO. High-oxidation degree GO had been synthesized using a modified Hummers' method. The synthesized GO was characterized using different analytical techniques including elemental analysis, Fourier transform infrared spectroscopy - universal attenuated total reflectance sensor (FTIR-UATR), Raman spectroscopy, and CHNSO elemental analysis. CHNSO analysis showed a high oxidation degree of GO represented by its oxygen content (50 wt.%). Then, ultrafiltration PSF membranes incorporating GO were fabricated using the phase inversion technique. The prepared membranes were characterized using scanning electron microscopy (SEM) and atomic force microscopy (AFM) and showed a clear effect of GO on PSF physical structure and morphology. The water contact angle of the membranes was measured and showed better hydrophilicity of GO membranes compared to pure PSF caused by the hydrophilic nature of GO. Separation properties of the prepared membranes were investigated using a cross-flow membrane system. Antifouling properties were studied using bovine serum albumin (BSA) and humic acid (HA) as model foulants. It has been found that GO-based membranes exhibit higher antifouling properties compared to pure PSF. When using BSA, the flux recovery ratio (FRR %) increased from 65.4 ± 0.9 % for pure PSF to 84.0 ± 1.0 % with a loading of 0.05 wt.% GO in PSF. When using HA as model foulant, FRR increased from 87.8 ± 0.6 % to 93.1 ± 1.1 % with 0.02 wt.% of GO in PSF. The pure water permeability (PWP) decreased with loadings of GO from 181.7 L.m⁻².h⁻¹.bar⁻¹ of pure PSF to 181.1, and 157.6 L.m⁻².h⁻¹.bar⁻¹ with 0.02 and 0.05 wt.% GO respectively. It can be concluded from the obtained results that incorporating low loading of GO could enhance the antifouling properties of PSF hence improving its lifetime and reuse.

Keywords: antifouling properties, GO based membranes, hydrophilicity, polysulfone, ultrafiltration

Procedia PDF Downloads 134

Authors: Aulanni’am Aulanni’am

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Incidence of Diabetes Mellitus (DM) progressively increasing it became a serious problem in Indonesia and it is a disease that government is priority to be addressed. The longer a person is suffering from diabetes the more likely to develop complications particularly diabetic patients who are not well maintained. Therefore, Incidence of Diabetes Mellitus needs to be done in the early diagnosis of pre-phase of the disease. In this pre-phase disease, already happening destruction of pancreatic beta cells and declining in beta cell function and the sign autoimmunity reactions associated with beta cell destruction. Type 1 DM is a multifactorial disease triggered by genetic and environmental factors, which leads to the destruction of pancreatic beta cells. Early marker of "beta cell autoreactivity" is the synthesis of autoantibodies against 65-kDa protein, which can be a molecule that can be detected early in the disease pathomechanism. The importance of early diagnosis of diabetic patients held in the phase of pre-disease is to determine the progression towards the onset of pancreatic beta cell destruction and take precautions. However, the price for this examination is very expensive ($ 150/ test), the anti-GAD65 abs examination cannot be carried out routinely in most or even in all laboratories in Indonesia. Therefore, production-based Rapid Test Recombinant Human Protein GAD65 with "Reverse Flow Immunchromatography Technique" in Indonesia is believed to reduce costs and improve the quality of care of patients with diabetes in Indonesia. Rapid Test Product innovation is very simple and suitable for screening and routine inspection of GAD65 autoantibodies. In the blood serum of patients with diabetes caused by autoimmunity, autoantibody-GAD65 is a major serologic marker to detect autoimmune reaction because their concentration level of stability.GAD65 autoantibodies can be found 10 years before clinical symptoms of diabetes. Early diagnosis is more focused to detect the presence autontibodi-GAD65 given specification and high sensitivity. Autoantibodies- GAD65 that circulates in the blood is a major indicator of the destruction of the islet cells of the pancreas. Results of research in collaboration with Biofarma has produced GAD65 autoantibodies based Rapid Test had conducted the soft launch of products and has been tested with the results of a sensitivity of 100 percent and a specificity between 90 and 96% compared with the gold standard (import product) which worked based on ELISA method.

Keywords: diabetes mellitus, GAD65 autoantibodies, rapid test, sensitivity, specificity

Procedia PDF Downloads 258

Authors: Waad Al Saleemi, Badriya Al Adawi, Zaaima Al Jabri, Sahim Al Ghafri, Jalila Al Hadhramia

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Objectives: Using standard lab methods, a positive blood culture requires a minimum of two days (two occasions of overnight incubation) to obtain a final identification (ID) and antimicrobial susceptibility results (AST) report. In this study, we aimed to evaluate the accuracy and precision of identification and antimicrobial susceptibility testing of an alternative method (direct method) that will reduce the turnaround time by 24 hours. This method involves the direct inoculation of positive blood culture broths into the Phoenix system using serum separation tubes (SST). Method: This prospective study included monomicrobial-positive blood cultures obtained from January 2022 to May 2023 in SQUH. Blood cultures containing a mixture of organisms, fungi, or anaerobic organisms were excluded from this study. The result of the new “direct method” under study was compared with the current “standard method” used in the lab. The accuracy and precision were evaluated for the ID and AST using Clinical and Laboratory Standards Institute (CLSI) recommendations. The categorical agreement, essential agreement, and the rates of very major errors (VME), major errors (ME), and minor errors (MIE) for both gram-negative and gram-positive bacteria were calculated. Passing criteria were set according to CLSI. Result: The results of ID and AST were available for a total of 158 isolates. Of 77 isolates of gram-negative bacteria, 71 (92%) were correctly identified at the species level. Of 70 isolates of gram-positive bacteria, 47(67%) isolates were correctly identified. For gram-negative bacteria, the essential agreement of the direct method was ≥92% when compared to the standard method, while the categorical agreement was ≥91% for all tested antibiotics. The precision of ID and AST were noted to be 100% for all tested isolates. For gram-positive bacteria, the essential agreement was >93%, while the categorical agreement was >92% for all tested antibiotics except moxifloxacin. Many antibiotics were noted to have an unacceptable higher rate of very major errors including penicillin, cotrimoxazole, clindamycin, ciprofloxacin, and moxifloxacin. However, no error was observed in the results of vancomycin, linezolid, and daptomycin. Conclusion: The direct method of ID and AST for positive blood cultures using SST is reliable for gram negative bacteria. It will significantly decrease the turnaround time and will facilitate antimicrobial stewardship.

Keywords: bloodstream infection, oman, direct ast, blood culture, rapid identification, antimicrobial susceptibility, phoenix, direct inoculation

Procedia PDF Downloads 51

Authors: Ludmila A. Gavriliuc, Jerry McLarty, Heather E. Kleiner, J. Michael Mathis

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Abstract -- Background: The citrus coumarine Auraptene (Aur) is an effective chemopreventive agent, as manifested in many models of diseases and cancer. Nuclear factor erythroid 2-related factor (Nrf2) is an important regulator of genes induced by oxidative stress, such as glutathione S-transferases, heme oxygenase-1, and peroxiredoxin 1, by activating the antioxidant response element (ARE). Genetic and biochemical evidence has demonstrated that glutathione (GSH) and glutathione-dependent enzymes, glutathione reductase (GR), glutathione peroxidases (GPs), glutathione S-transferases (GSTs) are responsible for the control of intracellular reduction-oxidation status and participate in cellular adaptation to oxidative stress. The effect of Aur on the activity of GR, GPs (Se-GP and Se-iGP), and content of GSH in the liver, kidney, and spleen is insufficiently explored. Aim: Our goal was the examination of the Aur influence on the redox-system of GSH in Nrf2 wild type and Nrf2 knockout mice via activation of Nrf2 and ARE. Methods: Twenty female mice, 10 Nrf2 wild-type (WT) and 10 Nrf2 (-/-) knockout (KO), were bred and genotyped for our study. The activity of GR, Se-GP, Se-iGP, GST, G6PD, CytP450 reductase, catalase (Cat), and content of GSH were analyzed in the liver, kidney, and spleen using Spectrophotometry methods. The results of the specific activity of enzymes and the amount of GSH were analyzed with ANOVA and Spearman statistical methods. Results: Aur (200 mg/kg) treatment induced hepatic GST, GR, Se-GP activity and inhibited their activity in the spleen of mice, most likely via activation of the ARE through Nrf2. Activation in kidney Se-GP and G6PD by Aur is also controlled, apparently through Nrf2. Results of the non-parametric Spearman correlation analysis indicated the strong positive correlation between GR and G6PD only in the liver in WT control mice (r=+0.972; p < 0.005) and in the kidney KO control mice (r=+0.958; p < 0.005). The observed low content of GSH in the liver of KO mice indicated an increase in its participation in the neutralization of toxic substances with the absence of induction of GSH-dependent enzymes, such as GST, GR, Se-GP, and Se-iGP. Activation of CytP450 in kidney and spleen and Cat in the liver in KO mice probably revealed another regulatory mechanism for these enzymes. Conclusion: Thereby, obtained results testify that Aur can modulate the activity of genes and antioxidant enzymatic redox-system of GSH, responsible for the control of intracellular reduction-oxidation status.

Keywords: auraptene, glutathione, GST, Nrf2

Procedia PDF Downloads 138

Authors: Mona Ghafouri-Azar, Sara Diamond, Jeremy Bowes, Grace Yuan, Aimee Burnett, Michelle Wyndham-West, Sara Wagner, Anand Pariyarath

Abstract:

Transportation is the fastest growing source of greenhouse gas emissions worldwide. In Canada, it is responsible for 23% of total CO2emissions from fuel combustion, and emissions from the transportation sector are the second largest source of emissions after the oil and gas sector. Currently, most Canadian public transportation systems rely on buses that operateon fossil fuels.Canada is currently investing billions of dollars to replacediesel buses with electric busesas this isperceived to have a significant impact on climate mitigation. This paper focuses on the possible impacts of zero emission buses (ZEB) on sustainable development, considering three dimensions of sustainability; environmental quality, economic growth, and social development.A sustainable transportation system is one that is safe, affordable, accessible, efficient, and resilient and that contributes minimal emissions of carbon and other pollutants.To enable implementation of these goals, relevant indicators were selected and defined that measure progress towards a sustainable transportation system. These were drawn from Canadian and international examples. Studies compare different European cities in terms of development, sustainability, and infrastructures, by using transport performance indicators. A Normalized Transport Sustainability index measures and compares policies in different urban areas and allows fine-tuning of policies. Analysts use a number ofmethods for sustainable analysis, like cost-benefit analysis (CBA) toassess economic benefit, life-cycle assessment (LCA) to assess social, economic, and environment factors and goals, and multi-criteria decision making (MCDM) analysis which can comparediffering stakeholder preferences.A multi criteria decision making approach is an appropriate methodology to plan and evaluate sustainable transit development and to provide insights and meaningful information for decision makers and transit agencies. It is essential to develop a system thataggregates specific discrete indices to assess the sustainability of transportation systems.Theseprioritize indicators appropriate for the differentCanadian transit system agencies and theirpreferences and requirements. This studywill develop an integrating index that alliesexistingdiscrete indexes to supporta reliable comparison between the current transportation system (diesel buses) and the new ZEB system emerging in Canada. As a first step, theindexes for each category are selected, and the index matrix constructed. Second, the selected indicators arenormalized to remove anyinconsistency between them. Next, the normalized matrix isweighted based on the relative importance of each index to the main domains of sustainability using the analytical hierarchy process (AHP) method. This is accomplished through expert judgement around the relative importance of different attributes with respect to the goals through apairwise comparison matrix. The considerationof multiple environmental, economic, and social factors (including equity and health) is integrated intoa sustainable transit planning index (STPI) which supportsrealistic ZEB implementation in Canada and beyond and is useful to different stakeholders, agencies, and ministries.

Keywords: zero emission buses, sustainability, sustainable transit, transportation, analytical hierarchy process, environment, economy, social

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