Search results for: fire assay

Authors: Amjad I. Aqib, Shanza R. Khan, Tanveer Ahmad, Syed A. R. Shah, Muhammad A. Naseer, Muhammad Shoaib, Iqra Sarwar, Muhammad F. A. Kulyar, Zeeshan A. Bhutta, Mumtaz A. Khan, Mahboob Ali, Khadija Yasmeen

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The current study focused on resistance modulation of dairy linked epidemic mec A positive S. aureus for resistance modulation by plant extract (Eucalyptus globolus, Calotropis procera), NSAIDs, and star like microparticles. Zinc oxide {ZnO}c and {Zn (OH)₂} microparticles were synthesized by solvothermal method and characterized by calcination, X-ray diffraction (XRD), and scanning electron microscope (SEM). Plant extracts were prepared by the Soxhlet extraction method. The study found 34% of subclinical samples (n=200) positive for S. aureus from dairy milk having significant (p < 0.05) association of assumed risk factors with pathogen. The antimicrobial assay showed 55, 42, 41, and 41% of S. aureus resistant to oxacillin, ciprofloxacin, streptomycin, and enoxacin. Amoxicillin showed the highest percentage of increase in zone of inhibitions (ZOI) at 100mg of Calotropis procera extract (31.29%) followed by 1mg/mL (28.91%) and 10mg/mL (21.68%) of Eucalyptus globolus. Amoxicillin increased ZOI by 42.85, 37.32, 29.05, and 22.78% in combination with 500 ug/ml with each of diclofenac, aspirin, ibuprofen, and meloxicam, respectively. Fractional inhibitory concentration indices (FICIs) showed synergism of amoxicillin with diclofenac and aspirin and indifferent synergy with ibuprofen and meloxicam. The preliminary in vitro finding of combination of microparticles with amoxicillin proved to be synergistic, giving rise to 26.74% and 14.85% increase in ZOI of amoxicillin in combination with zinc oxide and zinc hydroxide, respectively. The modulated antimicrobial resistance incurred by NSAIDs, plant extracts, and microparticles against pathogenic S. aureus invite immediate attention to probe alternative antimicrobial sources.

Keywords: antimicrobial resistance, dairy milk, nanoparticles, NSIDs, plant extracts, resistance modulation, S. aureus

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Authors: Rosemary Anibogwu, Kavita Sharma, Karl De Jesus

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Sagebrush is an abundant and naturally occurring plant in the Intermountain West region of the United States. The plant contains an array of bioactive compounds such as flavonoids, terpenoids, sterols, and phenolic acids. It is important to identify and characterize these compounds because Native Americans use sagebrush as herbal medicine. These compounds are also utilized for preventing infection in wounds, treating headaches and colds, and possess antitumor properties. This research is an exploratory study on the sesquiterpene present in the leaves of sagebrush. The leaf foliage was extracted with 100 % chloroform and 100 % methanol. The percentage yield for the crude was considerably higher in chloroform. The Thin Layer Chromatography (TLC) analysis of the crude extracted unveiled a brown band at Rf = 0.25 and a dark brown band at Rf = 0.74, along with three unknown faint bands the 254 nm UV lamp. Furthermore, the two distinct brown (Achillin) and dark brown band (Hydroxyachillin) in TLC were further utilized in the isolation of pure compounds with column chromatography. The structures of Achillin and Hydroxyachillin were elucidated based on extensive spectroscopic analysis, including TLC, High-Performance Liquid Chromatography (HPLC), 1D- and 2D-Nuclear Magnetic Resonance (NMR), and Mass Spectroscopy (MS). The antioxidant activities of crude extract and three pure compounds were evaluated in terms of their peroxyl radical scavenging by Ferric Reducing Ability of Plasma (FRAP) and 1,1-Diphenyl-2-picryl-hydrazyl (DPPH) methods. The crude extract showed the antioxidant activity of 18.99 ± 0.51 µmol TEg -1 FW for FRAP and 11.59 ± 0.38 µmol TEg -1 FW for DPPH. The activities of Achillin, Hydroxyachillin, and Quercetagetin trimethyl ether were 13.03, 15.90 and 14.02 µmol TEg -1 FW respectively for the FRAP assay. The three purified compounds have been submitted to the National Cancer Institute 60 cancer cell line for further study.

Keywords: HPLC, nuclear magnetic resonance spectroscopy, sagebrush, sesquiterpene lactones

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Authors: A. F. Agboola, B. R. O. Omidiwura, A. Oyeyemi, E. A. Iyayi, A. S. Adelani

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Dietary cholesterol has elicited the most public interest as it relates with coronary heart disease. Thus, humans have been paying more attention to health, thereby reducing consumption of cholesterol enriched food. Egg is considered as one of the major sources of human dietary cholesterol. However, an alternative way to reduce the potential cholesterolemic effect of eggs is to modify the fatty acid composition of the yolk. The effect of palm oil (PO), soybean oil (SO), sesame seed oil (SSO) and fish oil (FO) supplementation in the diets of layers on egg yolk fatty acid, cholesterol, egg production and egg quality parameters were evaluated in a 42-day feeding trial. One hundred and five Isa Brown laying hens of 34 weeks of age were randomly distributed into seven groups of five replicates and three birds per replicate in a completely randomized design. Seven corn-soybean basal diets (BD) were formulated: BD+No oil (T1), BD+1.5% PO (T2), BD+1.5% SO (T3), BD+1.5% SSO (T4), BD+1.5% FO (T5), BD+0.75% SO+0.75% FO (T6) and BD+0.75% SSO+0.75% FO (T7). Five eggs were randomly sampled at day 42 from each replicate to assay for the cholesterol, fatty acid profile of egg yolk and egg quality assessment. Results showed that there were no significant (P>0.05) differences observed in production performance, egg cholesterol and egg quality parameters except for yolk height, albumen height, yolk index, egg shape index, haugh unit, and yolk colour. There were no significant differences (P>0.05) observed in total cholesterol, high density lipoprotein and low density lipoprotein levels of egg yolk across the treatments. However, diets had effect (P<0.05) on TAG (triacylglycerol) and VLDL (very low density lipoprotein) of the egg yolk. The highest TAG (603.78 mg/dl) and VLDL values (120.76 mg/dl) were recorded in eggs of hens on T4 (1.5% sesame seed oil) and was similar to those on T3 (1.5% soybean oil), T5 (1.5% fish oil) and T6 (0.75% soybean oil + 0.75% fish oil). However, results revealed a significant (P<0.05) variations on eggs’ summation of polyunsaturated fatty acid (PUFA). In conclusion, it is suggested that dietary oils could be included in layers’ diets to produce designer eggs low in cholesterol and high in PUFA especially omega-3 fatty acids.

Keywords: dietary oils, egg cholesterol, egg fatty acid profile, egg quality parameters

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Authors: Laïd Boukraâ

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Complementary and Alternative Medicine is an inclusive term that describes treatments, therapies, and modalities that are not accepted as components of mainstream education or practice, but that are performed on patients by some practitioners. While these treatments and therapies often form part of post-graduate education, study and writing, they are generally viewed as alternatives or complementary to more universally accepted treatments. Ancient civilizations used bee products and medicinal plants, but modern civilization and ‘education’ have seriously lessened our natural instinctive ability and capability. Despite the fact that the modern Western establishment appears to like to relegate apitherapy and aromatherapy to the status of 'folklore' or 'old wives' tales', they contain a vast spread of pharmacologically-active ingredients and each one has its own unique combination and properties. They are classified in modern herbal medicine according to their spheres of action. Bee products and medicinal plants are well-known natural product for their healing properties and their increasing popularity recently as they are widely used in wound healing. Honey not only has antibacterial properties which can help as an antibacterial agent but also has chemical properties which may further help in the wound healing process. A formulation with honey as its main component was produced into a honey gel. This new formulation has enhanced texture and is more user friendly for usage as well. This new formulation would be better than other formulas as it is hundred percent consisting of natural products and has been made into a better formulation. In vitro assay, animal model study and clinical trials have shown the effectiveness of LEADERMAX for the treatment of diabetic foot, burns, leg ulcer and bed sores. This one hundred percent natural product could be the best alternative to conventional products for wound and burn management. The advantages of the formulation are: 100% natural, affordable, easy to use, strong power of absorption, dry surface on the wound making a film, will not stick to the wound bed; helps relieve wound pain, inflammation, edema and bruising while improving comfort.

Keywords: bed sore bee products, burns, diabetic foot, medicinal plants, leg ulcer, wounds

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Authors: Bahaa Eldin A. Fouda, Khaled N. Yossef, Mohamed Elhosseny, Ahmed Lotfy, Mohamed Salama, Mohamed Sobh

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Developmental neurotoxicity (DNT) entails the toxic effects imparted by various chemicals on the brain during the early childhood period. As human brains are vulnerable during this period, various chemicals would have their maximum effects on brains during early childhood. Some toxicants have been confirmed to induce developmental toxic effects on CNS e.g. lead, however; most of the agents cannot be identified with certainty due the defective nature of predictive toxicology models used. A novel alternative method that can overcome most of the limitations of conventional techniques is the use of 3D neurospheres system. This in-vitro system can recapitulate most of the changes during the period of brain development making it an ideal model for predicting neurotoxic effects. In the present study, we verified the possible DNT of epoxomicin which is a naturally occurring selective proteasome inhibitor with anti-inflammatory activity. Rat neural progenitor cells were isolated from rat embryos (E14) extracted from placental tissue. The cortices were aseptically dissected out from the brains of the fetuses and the tissues were triturated by repeated passage through a fire-polished constricted Pasteur pipette. The dispersed tissues were allowed to settle for 3 min. The supernatant was, then, transferred to a fresh tube and centrifuged at 1,000 g for 5 min. The pellet was placed in Hank’s balanced salt solution cultured as free-floating neurospheres in proliferation medium. Two doses of epoxomicin (1µM and 10µM) were used in cultured neuropsheres for a period of 14 days. For proliferation analysis, spheres were cultured in proliferation medium. After 0, 4, 5, 11, and 14 days, sphere size was determined by software analyses. The diameter of each neurosphere was measured and exported to excel file further to statistical analysis. For viability analysis, trypsin-EDTA solution were added to neurospheres for 3 min to dissociate them into single cells suspension, then viability evaluated by the Trypan Blue exclusion test. Epoxomicin was found to affect proliferation and viability of neuropsheres, these effects were positively correlated to doses and progress of time. This study confirms the DNT effects of epoxomicin on 3D neurospheres model. The effects on proliferation suggest possible gross morphologic changes while the decrease in viability propose possible focal lesion on exposure to epoxomicin during early childhood.

Keywords: neural progentor cells, epoxomicin, neurosphere, medical and health sciences

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Authors: Akram Abi, Clarissa Müller, Hans-Joachim Jördening

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Laminaribiose is a beta-1,3-glycoside which is used in the medical field for the treatment of dermatitis and also can be used as a building block for new pharmaceutics. The conventional process of laminaribiose production is the uneconomical process of hydrolysis of laminarin extracted from natural polysaccharides of plant origin. A more economical approach however is attainable by enzymatically synthesis of laminaribiose via a reverse phosphorylase reaction catalyzed by laminaribiose phosphorylase (LP) from Euglena gracilis. Different cultivation methods of Euglena gracilis and the effect on LP production have been investigated. Buffered/unbuffered heterotrophic and mixotrophic cultivations of Euglena gracilis has been carried out. Changes of biomass and LP production, glucose level and pH, cell count and shape has been monitored in the course of time. The results obtained from experiments each in three repetitions, show that in the heterotrophic cultivation of Euglena gracilis not only more biomass is produced compared to mixotrophic cultivation, but also higher specific protein concentration is achieved. Furthermore, the LP activity test showed that the protein extracted from heterotrophically cultured cells has a higher LP activity. It was also observed that the cells develop in a distinctive different shape between these two cultures and have different length to width ratios. Taking the heterotrophic culture as the more efficient cultivation method in LP production, another comparative experiment between buffered and unbuffered heterothrophic culture was carried out that showed the unbuffered culture has advantages over the other one in respect of both LP production and resulting activity. A hetrotrophic cultivation of Euglena gracilis in a 5L bioreactor with controlled operating conditions showed a distinctive improvement of all the aspects of culture compared to the shaking flask cultivations. Biomass production was improved from 5 to more than 8 g/l (dry weight) which resulted in a specific protein concentration of 45 g/l in the heterotrophic cultivation in the bioreactor. In further attempts to improve LP production, different purification methods were tested and each method was checks through an activity assay. A laminaribiose yield of 35% was achieved which was by far the highest amount amongst different methods tested.

Keywords: euglena gracilis, heterotrophic culture, laminaribiose production, mixotrophic culture

Procedia PDF Downloads 365

Authors: Suveen Kumar

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Oral cancer (OC) is the sixth most death causing cancer in world which includes tumour of lips, floor of the mouth, tongue, palate, cheeks, sinuses, throat, etc. Conventionally, the techniques used for OC detection are toluidine blue staining, biopsy, liquid-based cytology, visual attachments, etc., however these are limited by their highly invasive nature, low sensitivity, time consumption, sophisticated instrument handling, sample processing and high cost. Therefore, we developed biosensing chips for non-invasive detection of OC via CYFRA-21-1 biomarker. CYFRA-21-1 (molecular weight: 40 kDa) is secreted in saliva of OC patients which is a non-invasive biological fluid with a cut-off value of 3.8 ng mL-1, above which the subjects will be suffering from oral cancer. Therefore, in first work, 3-aminopropyl triethoxy silane (APTES) functionalized zirconia (ZrO2) nanoparticles (APTES/nZrO2) were used to successfully detect CYFRA-21-1 in a linear detection range (LDR) of 2-16 ng mL-1 with sensitivity of 2.2 µA mL ng-1. Successively, APTES/nZrO2-RGO was employed to prevent agglomeration of ZrO2 by providing high surface area reduced graphene oxide (RGO) support and much wider LDR (2-22 ng mL-1) was obtained with remarkable limit of detection (LOD) as 0.12 ng mL-1. Further, APTES/nY2O3/ITO platform was used for oral cancer bioseneor development. The developed biosensor (BSA/anti-CYFRA-21-1/APTES/nY2O3/ITO) have wider LDR (0.01-50 ng mL-1) with remarkable limit of detection (LOD) as 0.01 ng mL-1. To improve the sensitivity of the biosensing platform, nanocomposite of yattria stabilized nanostructured zirconia-reduced graphene oxide (nYZR) based biosensor has been developed. The developed biosensing chip having ability to detect CYFRA-21-1 biomolecules in the range of 0.01-50 ng mL-1, LOD of 7.2 pg mL-1 with sensitivity of 200 µA mL ng-1. Further, the applicability of the fabricated biosensing chips were also checked through real sample (saliva) analysis of OC patients and the obtained results showed good correlation with the standard protein detection enzyme linked immunosorbent assay (ELISA) technique.

Keywords: non-invasive, oral cancer, nanomaterials, biosensor, biochip

Procedia PDF Downloads 127

Authors: Md. Soebur Rahman, Mahbuba Begum, Raquib Ahsan

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Composite column is a structural member that uses a combination of structural steel shapes, pipes or tubes with or without reinforcing steel bars and reinforced concrete to provide adequate load carrying capacity to sustain either axial compressive loads alone or a combination of axial loads and bending moments. Composite construction takes the advantages of the speed of construction, light weight and strength of steel, and the higher mass, stiffness, damping properties and economy of reinforced concrete. The most usual types of composite columns are the concrete filled steel tubes and the partially or fully encased steel profiles. Fully encased composite column (FEC) provides compressive strength, stability, stiffness, improved fire proofing and better corrosion protection. This paper reports experimental and numerical investigations of the behaviour of concrete encased steel composite columns subjected to short-term axial load. In this study, eleven short FEC columns with square shaped cross section were constructed and tested to examine the load-deflection behavior. The main variables in the test were considered as concrete compressive strength, cross sectional size and percentage of structural steel. A nonlinear 3-D finite element (FE) model has been developed to analyse the inelastic behaviour of steel, concrete, and longitudinal reinforcement as well as the effect of concrete confinement of the FEC columns. FE models have been validated against the current experimental study conduct in the laboratory and published experimental results under concentric load. It has been observed that FE model is able to predict the experimental behaviour of FEC columns under concentric gravity loads with good accuracy. Good agreement has been achieved between the complete experimental and the numerical load-deflection behaviour in this study. The capacities of each constituent of FEC columns such as structural steel, concrete and rebar's were also determined from the numerical study. Concrete is observed to provide around 57% of the total axial capacity of the column whereas the steel I-sections contributes to the rest of the capacity as well as ductility of the overall system. The nonlinear FE model developed in this study is also used to explore the effect of concrete strength and percentage of structural steel on the behaviour of FEC columns under concentric loads. The axial capacity of FEC columns has been found to increase significantly by increasing the strength of concrete.

Keywords: composite, columns, experimental, finite element, fully encased, strength

Procedia PDF Downloads 290

Authors: Tanakrit Doltanakarn

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Introduction: These days, cannabis is being accepted in many countries due to the fact that cannabis could be use in medical. The medical cannabis is used to treat and reduce the pain many diseases. For example, neuropathic pain, Parkinson, autism disorders, cancer pain reduce the adverse effect of chemotherapy, diabetes, and migraine. Active ingredients in cannabis that modulate patients' perceptions of their conditions include Δ9‐tetrahydrocannabinol (THC), cannabidiol (CBD), flavonoids, and terpenes. However, there is an adverse effect of cannabis, cardiovascular effects, psychosis, schizophrenia, mood disorder, and cognitive alternation. These effects are from the THC and CBD ingredients in the cannabis. The metabolize processes of delta-9 THC to 11-OH-delta 9 -THC (inactive form), THC were cause of adverse effects. Interestingly, the distributions of CYP2C9 gene (CYP2C9*2 and CYP2C9*3, poor metabolizer) that might affect incidences of adverse effects in patients who treated with medical cannabis. Objective: The aim of this study we want to investigate the association between genetic polymorphism of CYP2C9 frequency and Thai patients who treated with medical cannabis. Materials and Methods:We recruited sixty-five unrelated Thai patients from the College of Pharmacy, Rangsit University. DNA were extracted using Genomic DNA Mini Kit. Genotyping of CYP2C9*2 (430C>T, rs1799853) and CYP2C9*3 (1075A>C, rs1057910) were genotyped by the TaqMan Real-time PCR assay. Results: Among these 31 medicals cannabis-induced ADRs patients, they were diagnosed with 22 (33.85%) tachycardia and 3 (4.62%) arrhythmia. There were 34 (52.31%) medical cannabis-tolerant controls who were included in this study.40 (61.53%) Thai patients were female, and 25 (38.46%) were male, with median age of 57 (range 27 – 87) years. In this study, we found none of the medical cannabis-induced ADRs carried CYP2C9*2 variant along with medical cannabis-tolerant control group. CYP2C9*3 variant (intermediate metabolizer, IM) was found just only one of thirty-one (3.23%) in the medical cannabis-induced ADRs and two of thirty-fourth (5.88%) in the tolerant controls. Conclusions: Thus, the distribution of CYP2C9 alleles offer a comprehensive view of pharmacogenomics marker in Thai population that could be used as a reference for worldwide to investigate the pharmacogenomics application.

Keywords: medical cannabis, adverse effect, CYP2C9, thai patients

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Authors: Denise Schrama, Claudia Raposo, Pedro Rodrigues

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Background: Food allergies are conducted by a hypersensitive response of the immune system. These allergies are a global concern for the public health. Consumption of fish is increasing worldwide as it is a healthy meat with high nutritional value. Unfortunately, fish can cause adverse immune-mediate reactions, affecting part of the population with higher incidence in children. β-parvalbumin, a small, highly conserved stable, calcium or magnesium binding muscle protein is the main fish allergen. In fish-allergic patients, cross-reactivity between different fish species exist due to recognition of highly identical protein regions. Enolases, aldolases, or fish gelatin are other identified fish allergens in some fish species. With no available cure for fish allergies, clinical management is only based on an avoidance diet aiming at the total exclusion of offending food. Methods: Mediterranean fish (S. aurata and D. labrax) were fed specifically designed diets, enriched in components that target the expression or inactivation of parvalbumin (creatine and EDTA, respectively). After 90 days fish were sampled and biological tissues were excised. Proteomics was used to access fish allergens characterization and expression in muscle while IgE assays to confirm the lower allergenic potential are conducted in patients with history of fish allergies. Fish welfare and quality of flesh were established with biochemical, texture and sensorial analysis. Results: Fish welfare shows no major impact between diets. In case of creatine supplementation in D. labrax proteomic analysis show a slight decrease in parvalbumin expression. No accumulation of this compound was found in muscle. For EDTA supplementation in S. aurata IgE assay show a slight decrease in allergenicity when using sera of fish allergic patients. Conclusion: Supplementation with these two compounds seems to change slightly the allergenicity of the two mean Mediterranean species.

Keywords: fish allergies, fish nutrition, proteomics, aquaculture

Procedia PDF Downloads 156

Authors: Ji-Young Choi, Jungjin Kim, Sang-Sun Yun, Sangmee Ahn Jo

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Intracellular adhesion molecule1 (ICAM1) is a mediator of inflammation and involved in adhesion and transmigration of leukocytes to endothelial cells, resulting in enhancement of brain inflammation. We hypothesized that increase of ICAM1 expression in endothelial cells is an early step in the pathogenesis of brain diseases such as Alzheimer’s disease. Here, we report that ICAM1 expression is regulated by pro-inflammatory cytokine TNFa in human microvascular endothelial cell (HBMVEC). TNFa significantly increased ICAM1 mRNA and protein levels at the concentrations showing no cell toxicity. This increase was also shown in micro vessels of mouse brain 24 hours after treatment with TNFa (8 mg/kg, i.v). We then investigated the epigenetic mechanism involved in the induction of ICAM1 expression. Chromatin immunoprecipitation assay revealed that TNFa reduced methylation of histone3K9 (H3K9-2me) and histone3K27 (H3K27-3me), well-known modification as gene suppression, with in the ICAM1 promoter region. However, acetylation of H3K9 and H3K14, well-known modification as gene activation, was not changed by TNFa. Treatment of BIX01294, a specific inhibitor of histone methyltransferase G9a responsible for H3K9-2me, dramatically increased in ICAM1 mRNA and protein levels and overexpression of G9a gene suppressed TNFa-induced ICAM1 expression. In contrast, GSK126, an inhibitor of histone methyltransferase EZH2 responsible for H3K27-3me and valproic acid, an inhibitor of histone deacetylase (HDAC) did not affect ICAM1 expression. These results suggested that histone3 methylation is involved in ICAM1 repression. Moreover, TNFa or BIX01294-induced ICAM induction resulted in both enhancements in adhesion and transmigration of leukocyte on endothelial cell. This study demonstrates that TNFa upregulates ICAM1 expression through H3K9-2me and H3K27-3me within the ICAM1 promoter region, in which G9a is likely to play a pivotal role in ICAM1 transcription. Our study provides a novel mechanism for ICAM1 transcription regulation in HBMVEC.

Keywords: ICAM1, TNFa, HBMVEC, H3K9-2me

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Authors: Małgorzata Tomczyńska, Joanna Saluk-Bijak

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Background: Graves’ disease is a frequent organ-specific autoimmune thyroid disease, which characterized by the presence of different kind autoantibodies, that, in most cases, act as agonists of the thyrotropin receptor, leading to hyperthyroidism. Role of platelets and lymphocytes can be modulated in the pathophysiology of thyroid autoimmune diseases. Interference in the physiology of platelets can lead to enhanced activity of these cells. Activated platelets can bind to circulating lymphocytes and to affect lymphocyte adhesion. Platelets and lymphocytes can regulate mutual functions. Therefore, the activation of T lymphocytes, as well as blood platelets, is associated with the development of inflammation and oxidative stress within the target tissue. The present study was performed to investigate a platelet-lymphocyte relation by assessing the degree of their mutual aggregation in whole blood of patients with Graves’ disease. Also, the purpose of this study was to examine the impact of platelet interaction on lymphocyte migration capacity. Methods: 30 patients with Graves’ disease were recruited in the study. The matched 30 healthy subjects were served as the control group. Immunophenotyping of lymphocytes was carried out by flow cytometry method. A CytoSelect™ Cell Migration Assay Kit was used to evaluate lymphocyte migration and adhesion to blood platelets. Visual assessment of lymphocyte-platelet aggregate morphology was done using confocal microscope after magnetic cell isolation by Miltenyi Biotec. Results: The migration and functional responses of lymphocytes to blood platelets were greater in the group of Graves’ disease patients compared with healthy controls. The group of Graves’ disease patients exhibited a reduced T lymphocyte and a higher B cell count compared with controls. Based on microscopic analysis, more platelet-lymphocyte aggregates were found in patients than in control. Conclusions: Studies have shown that in Graves' disease, lymphocytes show increased platelet affinity, more strongly migrating toward them, and forming mutual cellular conglomerates. This may be due to the increased activation of blood platelets in this disease.

Keywords: blood platelets, cell migration, Graves’ disease, lymphocytes, lymphocyte-platelet aggregates

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Authors: Said Abdirasak Abidrahman, Ibrahim Mohamed

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Background: Urinary Tract Infections (UTIs) are the commonest infections described among diabetes mellitus patients. More often, empirical antimicrobial therapy is initiated before the laboratory results are made available with minimal treatment success. The knowledge of the etiology and antibiotic susceptibility patterns of the organisms causing urinary tract infections among diabetes mellitus patients remains scarce, despite its vitality. This study sought to determine the prevalence, bacteria species, and drug susceptibility patterns of common causes of urinary tract infections among diabetes mellitus patients attending Bosaso health centers. Materials and methods: We conducted a cross-sectional study involving adult diabetic patients at Bosaso health centers between the months of May and July 2020. Laboratory assay of mid-stream urine samples was done to isolate bacteria causes of UTIs. These were biochemically identified using Gram stain, Kligler iron agar (KIA), Indole test, citrate, urea, coagulase, catalase, motility agar, and lysine iron agar. Their antibiotic susceptibility pattern for the isolated organisms was made for Ampicillin 10μg, Ciprofloxacin 5μg, Cotrimoxazole 25μg, Gentamycin 10μg, Ceftriaxone 10μg, and determined using the Kirby Bauer Disc Diffusion method. Results: Of 177 participants, 69 (39.0%) were males and 108 (61.0%) were females. Their mean age was 33.1 years (range; 18-67 years). Of these, 14.7% (26/177) of the samples revealed significant growth (>= 105 CFU/mL) giving a prevalence of 14.9 % (95% CI: 10.6 to 16.3). The organisms isolated were Escherichia coli -50% (N=13), Klebsiella pneumonia 30.8% (N=8), Staphylococcus aureus 15.4% (N=4), and unidentified organism 3.8% (N=1), and these were associated with such socio-demographic factors like history of catheterization and sexual activity. Antibiotic susceptibility to the commonly used agents for treating UTIs indicated higher sensitivity to Gentamicin and Ceftriaxone.

Keywords: antimicrobials, bacteria, urinary tract infections, diabetes

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Authors: Yemima Dani Riani, Anggraini Barlian

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Green turtle (Chelonia mydas) is one of well known long-lived turtle. Its age can reach 100 years old. Senescence in green turtle is an interesting process to study because until now no clear explanation has been established about senescence at cellular or molecular level in this species. Since 1999, green turtle announced as an endangered species. Hence, establishment of fibroblast skin cell culture of green turtle may be material for future study of senescence. One common marker used for detecting senescence is telomere shortening. Reduced telomerase activity, the reverse transcriptase enzyme which adds TTAGGG DNA sequence to telomere end, may also cause senescence. The purpose of this research are establish and identify green turtle fibroblast skin cell culture and also compare telomere length and telomerase activity from passage 5 and 14. Primary cell culture made with primary explant method then cultured in Leibovitz-15 (Sigma) supplemented by 10% Fetal Bovine Serum (Sigma) and 100 U/mL Penicillin/Streptomycin (Sigma) at 30 ± 1oC. Cells identified with Rabbit Anti-Vimentin Polyclonal Antibody (Abcam) and Goat Polyclonal Antibody (Abcam) using confocal microscope (Zeiss LSM 170). Telomere length obtained using TeloTAGGG Telomere Length Assay (Roche) while telomerase activity obtained using TeloTAGGG Telomerase PCR ElisaPlus (Roche). Primary cell culture from green turtle skin had fibroblastic morphology and immunocytochemistry test with vimentin antibody proved the culture was fibroblast cell. Measurement of telomere length and telomerase activity showed that telomere length and telomerase activity of passage 14 was greater than passage 5. However, based on morphology, green turtle fibroblast skin cell culture showed senescent morphology. Based on the analysis of telomere length and telomerase activity, suspected fibroblast skin cell culture of green turtles is not undergo aging through telomere shortening.

Keywords: cell culture, chelonia mydas, telomerase, telomere, senescence

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Authors: Z. Wochyński, K. A. Sobiech, Z. Kobos

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To C-Reactive Proteins include ferritin, transferrin, and ceruloplasmin- metalloproteins. The study aimed at assessing an effect of training on the Special Aerial Gymnastics Instruments (SAGI) on changes of serum ferritin, transferrin, and ceruloplasmin and cadets’ physical fitness in comparison with a control group. Fifty-five cadets in the mean age 20 years were included into this study. They were divided into two groups: Group A (N=41) trained on SAGI and Group B (N=14) trained according the standard program of physical education (control group). In both groups, blood was a material for assays. Samples were collected twice before and after training at the start of the program (training I), during (training II), and after education program completion (training III). Commercially available kits were used to assay blood serum ferritin, transferrin, and ceruloplasmin. Cadets’ physical fitness was evaluated with exercise tests before and after education program completion. In Group A, serum post-exercise ferritin decreased statistically insignificantly in training I and II and increased in training III in comparison with pre-exercise values. In Group B, post-exercise serum ferritin decreased statistically insignificantly in training I and III and significantly increased in training II in comparison with the pre-exercise values. In Group A, serum transferrin decreased statistically insignificantly in training I, and significantly increased in training II, whereas in training III it increased insignificantly in comparison with pre-exercise values. In Group B, post-exercise serum transferrin increased statistically significantly in training I, II, and III in comparison with pre-exercise values. I n Group A, serum ceruloplasmin decreased in all three series in comparison with pre-exercise values. In Group B, serum ceruloplasmin increased significantly in training II. It was showed that the training on SAGI significantly decreased serum ceruloplasmin in Group A in all three series of assays and did not produce significant changes in serum ferritin also was showed significant increase in serum transferrin.

Keywords: special aerial gymnastics instruments, ferritin, ceruloplasmin, transferrin

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Authors: Marina G. Galitskaya, Svetlana G. Makarova, Andrey P. Fisenko.

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Children on medication-induced immunosuppression are at high risk of developing severe course infectious diseases. Therefore, preventive vaccination is especially important for these children. However, due to the immunosuppressive effects of treatment for the underlying disease, the effectiveness of vaccination may decrease below the protective level. In a multidisciplinary children's medical center, post-vaccination immunity was studied in 79 children aged 4-17 years. The children were divided into 2 groups: Group 1 (38 children) with kidney pathology (Nephrotic Syndrome) and Group 2 (41 children) with inflammatory bowel diseases (Ulcerative Colitis, Crohn's Disease). Both groups of children were vaccinated according to the national vaccination calendar and received immunosuppressive therapy (prednisolone, methotrexate, cyclosporine, and other drugs) for at least 1 year. Using the enzyme-linked immunosorbent assay method, specific IgG antibodies to vaccine-preventable infections were determined: measles, rubella, mumps, diphtheria, pertussis, tetanus, and hepatitis B. The study showed the percentage of children with positive IgG values for vaccine-preventable infections. The highest percentage of children had protective antibody levels to measles (84.2% in children with nephrotic syndrome and 92.6% in those with inflammatory bowel disease) and rubella (71% and 80.4%, respectively). The lowest percentage of children with protective antibodies was for hepatitis B (5.2% and 29.2% respectively). Antibodies to mumps, diphtheria, pertussis, and tetanus were found not in all children (from 39,4% to 82,9%). The remaining percentage of children did not have detectable IgG antibodies to vaccine-preventable infections. Not all children, despite the previous vaccination, preserved antibodies to vaccine-controlled infections and remained unprotected by specific IgG antibodies. The issue of a booster vaccine dose should be considered in children without contraindications to vaccination. Children receiving long-term immunosuppressive therapy require an individual vaccination approach, including a specific definition of the performed vaccination.

Keywords: immunosuppressive therapy, inflammatory bowel diseases, nephrotic syndrome, post-vaccination immunity, specific antibodies, vaccine-preventable infections.

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Authors: Aliya Sekenova, Vyacheslav Ogay

Abstract:

The derivation of human induced pluripotent stem cells (iPSCs) from somatic cells by direct reprogramming opens wide perspectives in the regenerative medicine. It means the possibility to develop the personal and, consequently, any immunologically compatible cells for applications in cell-based therapy. The purpose of our study was to develop the technology for the production of NK cells from T cell-derived induced pluripotent stem cells (TiPSCs) for subsequent application in adoptive cancer immunotherapy. Methods: In this study iPSCs were derived from peripheral blood T cells using Sendai virus vectors expressing Oct4, Sox2, Klf4 and c-Myc. Pluripotent characteristics of TiPSCs were examined and confirmed with alkaline phosphatase staining, immunocytochemistry and RT-PCR analysis. For NK cell differentiation, embryoid bodies (EB) formed from (TiPSCs) were cultured in xenogeneic serum-free medium containing human serum, IL-3, IL-7, IL-15, SCF, FLT3L without using M210-B4 and AFT-024 stromal feeder cells. After differentiation, NK cells were characterized with immunofluorescence analysis, flow cytometry and cytotoxicity assay. Results: Here, we for the first time demonstrate that TiPSCs can effectively differentiate into functionally active NK cells without M210-B4 and AFT-024 xenogeneic stroma cells. Immunofluorescence and flow cytometry analysis showed that EB-derived cells can differentiate into a homogeneous population of NK cell expressing high levels of CD56, CD45 and CD16 specific markers. Moreover, these cells significantly express killing activation receptors such as NKp44 and NKp46. In the comparative analysis, we observed that NK cells derived using feeder-free culture system have more high killing activity against K-562 tumor cells, than NK cells derived by feeder-dependent method. Thus, we think that our obtained data will be useful for the development of large-scale production of NK cells for translation into cancer immunotherapy.

Keywords: induced pluripotent stem cells, NK cells, T cells, cell diffentiation, feeder cell-free culture system

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Authors: Vasilis Andriopoulos, Maria D. Gkioni, Elena Koutra, Savvas G. Mastropetros, Fotini N. Lamari, Sofia Hatziantoniou, Michalis Kornaros

Abstract:

In the present study, the antioxidant activity of aqueous and methanolic extracts of Chlorella minutissima, Dunaliella salina, Isochrysis galbana, Nannochloropsis oculata and Tisohrysis lutea, as well as the proximate composition and fatty acid profile were evaluated, with the aim to select species suitable for co-production of antioxidants and aquaculture feed. Batch cultivation was performed at 25oC in a modified f/2 medium under continuous illumination and aeration with ambient air. Biomass was collected via centrifugation and extracted first with H2O and subsequently with methanol at two growth phases (early and late stationary). Total phenolic content and antioxidant and reducing activity of the extracts were evaluated. The highest phenolic content was found in the methanolic extract of C. minutissima at the early stationary phase (9.04±0.68 mg Gallic Acid Equivalent g-1 dry weight), and the aqueous extract of D. salina at the late stationary phase (8.78±1.49 mg Gallic Acid Equivalent g-1 Dry weight). Antioxidant activity, measured as 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, and Ferric reducing antioxidant power assay of methanolic extracts were comparable to the literature and correlated to Total phenolic content and Chlorophyll content of the biomass. No such correlation was found in the aqueous extracts. N. oculata and T. lutea were high in protein (39.88±1.72% Dry weight and 43.30±1.33% Dry weight, respectively) and carotenoids (0.64±0.13% and 0.92±0.02%, respectively). Additionally, they presented high eicosapentaenoic acid and docosahexaenoic acid levels (33.74±9.98 mg eicosapentaenoic acid g-1 DW and 31.31±2.92 mg docosahexaenoic acid g-1 dry weight, respectively). N. oculata and T. lutea are promising candidates for the co-production of antioxidants and aquaculture feed, while C. minutissima and D. salina showed promise due to their higher antioxidant content.

Keywords: aquaculture fee, antioxidant activity, fatty acids, microalgae, total phenolic content

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Authors: Howaida I. Abd-Alla, Omnia Kutkat, Yassmin Moatasim, Magda T. Ibrahim, Marwa A. Mostafa, Mohamed GabAllah, Mounir M. El-Safty

Abstract:

Artemisia judaica (known shih-balady), Azadirachta indica and Sophora tomentosa (known yellow necklace pod) are members of available medicinal plants well-known for their traditional medical use in Egypt which suggests that they probably harbor broad-spectrum antiviral, immunostimulatory and anti-inflammatory functions. Their ethyl acetate-dichloromethane (1:1, v/v) extracts were evaluated for the potential anti-Middle East respiratory syndrome-related coronavirus (anti-MERS-CoV) activity. Their cytotoxic activity was tested in Vero-E6 cells using 3-(4,-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method with minor modification. The plot of percentage cytotoxicity for each extract concentration has calculated the concentration which exhibited 50% cytotoxic concentration (TC50). A plaque reduction assay was employed using safe dose of extract to evaluate its effect on virus propagation. The highest inhibition percentage was recorded for the yellow necklace pod, followed by Shih-balady. The possible mode of action of virus inhibition was studied at three different levels viral replication, viral adsorption and virucidal activity. The necklace pod leaves have induced virucidal effects and direct effects on the replication of virus. Phytochemical investigation of the promising necklace pod led to the isolation and structure determination of nine compounds. The structure of each compound was determined by a variety of spectroscopic methods. Compounds 4-O-methyl sorbitol 1, 8-methoxy daidzin 6 and 6-methoxy apigenin-7-O-β-D-glucopyranoside 8 were isolated for the first time from the Sophora genus and the other six compounds were the first time that they were isolated from this species according to available works of literature. Generally, the highest anti-CoV 2 activity of S. tomentosa was associated with the crude ethanolic extract, indicating the possibility of synergy among the antiviral phytochemical constituents (1-9).

Keywords: coronavirus, MERS-CoV, mode of action, necklace pod, shih-balady

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Authors: Anna Pick Kiong Ling, Jia May Chin, Rhun Yian Koh, Ying Pei Wong

Abstract:

Background: Uncontrolled inflammation may cause serious inflammatory diseases if left untreated. Non-steroidal anti-inflammatory drug (NSAIDs) is commonly used to inhibit pro-inflammatory enzymes, thus, reduce inflammation. However, long term administration of NSAIDs leads to various complications. Medicinal plants are getting more attention as it is believed to be more compatible with human body. One of them is a flavonoid-containing medicinal plants, Strobilanthes crispus which has been traditionally claimed to possess anti-inflammatory and antioxidant activities. Nevertheless, its anti-inflammatory activities are yet to be scientifically documented. Objectives: This study aimed to examine the anti-inflammatory activity of S. crispus by investigating its effects on intracellular oxidative stress and prostaglandin E₂ (PGE₂) levels. Materials and Methods: In this study, the Maximum Non-toxic Dose (MNTD) of methanol extract of both leaves and stems of S. crispus was first determined using 3-(4,5-dimethylthiazolyl-2)-2,5-diphenytetrazolium Bromide (MTT) assay. The effects of S. crispus extracts at MNTD and half MNTD (½MNTD) on intracellular ROS as well as PGE₂ levels in 1.0 µg/mL LPS-stimulated RAW 264.7 macrophages were then be measured using DCFH-DA and a competitive enzyme immunoassay kit, respectively. Results: The MNTD of leaf extract was determined as 700µg/mL while for stem was as low as 1.4µg/mL. When LPS-stimulated RAW 264.7 macrophages were subjected to the MNTD of S. crispus leaf extract, both intracellular ROS and PGE₂ levels were significantly reduced. In contrast, stem extract at both MNTD and ½MNTD did not significantly reduce the PGE₂ level, but significantly increased the intracellular ROS level. Conclusion: The methanol leaf extract of S. crispus may possess anti-inflammatory properties as it is able to significantly reduce the intracellular ROS and PGE₂ levels of LPS-stimulated cells. Nevertheless, further studies such as investigating the interleukin, nitric oxide and cytokine tumor necrosis factor-α (TNFα) levels has to be conducted to further confirm the anti-inflammatory properties of S. crispus.

Keywords: anti-inflammatory, natural products, prostaglandin E2, reactive oxygen species

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Authors: Zeb Hussain, Muhammad Asif Qureshi, Shaheen Sharafat

Abstract:

Background: Measles is a contagious viral infection common in childhood. Vaccination against measles is included in the expanded program of immunization (EPI). However, and alarmingly, a high mortality rate is observed due to measles infection in Pakistan. Moreover a recent outbreak of measles in various areas of Pakistan further highlights the problem. It is therefore important to investigate measles specific IgG (antibody) levels in our population. Objective: To quantify measles specific IgG antibodies amongst vaccinated children in district Qamber Shahdadkot, Sindh. Methodology: This cross-sectional study was conducted at the Microbiology section of the Dow-Diagnostic-Research-and-Reference-Laboratory (DDRRL), DUHS after Institutional Review Board approval (IRB-516/DUHS/-14) during August-December-2014. A total of 173 participants (residents of district Qamber Shahdadkot, Sindh) aged between 1-5 years were recruited in the study. Blood samples were collected as per standard phlebotomy guidelines. Blood was stored at 4 °C overnight. Samples were subsequently spun at a speed of 10000rpm to separate sera, which were divided into small aliquots to be frozen at -20 °C. Frozen sera were transported to the DDRRL on dry ice. Measles specific IgG (antibody) titers were quantified using enzyme linked immunosorbant assay (ELISA). Results: Blood was collected from a total of 173 individuals ranging between 1-5 years of age. Of these, a total of 88 participants were males and 85 were females. Of the 173 investigated samples, only 53 (30.6%) showed protective IgG titers against measles while 120 (69%) were sero-negative. Measles specific IgG antibodies titers were higher in female participants compared to the males. Conclusion: Our data demonstrate that a substantial percentage of vaccinated children in district Qamber-Shahdadkot did not have protective antibody titres against measles. It is therefore extremely important to investigate measles specific IgG levels in various parts of Pakistan in order to implement appropriate protective measures.

Keywords: sero-surveillance, measles, vaccinated children, Pakistan

Procedia PDF Downloads 331

Authors: Stéfani T. A. Dantas, Laura T. S. Takume, Bruna F. Rossi, Érika R. Bonsaglia, Ivana G. Castilho, José C. F. Pantoja, Ary Fernandes Júnior, Juliano L. Gonçalves, Marcos V. Santos, Rinaldo A. Mota, Vera L. M. Rall

Abstract:

Staphylococcus aureus is one of the main pathogens causing contagious bovine mastitis, being more frequently isolated from subclinical form, although the clinical form also occurs. Clinical mastitis cause visual signs, such as swelling, fever, hardening of the mammary gland, or any change in the characteristics of the milk. Considering the subclinical type, there are no visible signs in the animal nor changes in the milk. S. aureus has many important virulence factors for the establishment of its pathogenicity in animals, such as enterotoxins, which are also responsible for foodborne poisoning. Our objective is to perform a comparative analysis between 103 isolates of S. aureus, obtained from the milk of cows with clinical mastitis and 103 more, from subclinical type, in relation to the presence of these enterotoxins and verify if their presence plays an important role in the signs of illness. We will investigate all enterotoxins described till now, such as sea-see, seg-sez, sel26, sel 27, se01, and se02 (This study was approved by the Sao Paulo State University Animal Use Ethics Committee, No. 0136/2017). For the PCR assay, we used Illustra Bacteria Mini Spin Kit for bacterial DNA. At this moment, we have already tested sea-see, seg-ser, sew, and sex, and the results have already been submitted to Fisher Exact Probability Test or Chi-square Test. Considering the isolates obtained from clinical mastitis, the most frequent enterotoxins were selw (99%), selx (78%) and selh (50.5%), and sec, see, sej, sell, selp,and ser were absent. Among the subclinics, selw (82.5%) selm (15.5%) and selx (14.6%) were the most frequent, and sea-see, seg, sei-sel, sem-ser were absent. We have already observed statistically significant differences for seb, seg, seh, sei, selo, selu, selw and selx. Other interesting results were the low number of genes in each isolate from subclinical mastitis [0 genes: 14 (13.6%); 1 gene: 55 (53.4%); 2 genes: 33 (32%) or 3: 1 (0.97%)] compared to clinical isolates [1 gene: 5 (4.9%); 2 genes: 29 (28.1%); 3 genes: 38 (36.9%); 4 genes: 14 (13.6%); 5 genes: 5 (4.9%); 6 genes: 4 (3.9%); 7 genes: 5 (4.9%); 8 genes: 2 (1.9%) and 9 genes: 1 (1%)]. Based on these results, we can conclude that enterotoxins indeed play an important role in clinical signs in cattle with mastitis.

Keywords: mastitis, S. aureus, PCR, staphylococcal enterotoxin

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Authors: Mohammed Alasel, Michael Keusgen

Abstract:

Gold is a noble metal; in its nano-scale level (e.g. spherical nanoparticles), the conduction electrons are triggered to collectively oscillate with a resonant frequency when certain wavelengths of electromagnetic radiation interact with its surface; this phenomenon is known as surface plasmon resonance (SPR). SPR is responsible for giving the gold nanoparticles its intense red color depending mainly on its size, shape and distance between nanoparticles. A decreased distance between gold nanoparticles results in aggregation of them causing a change in color from red to blue. This aggregation enables gold nanoparticles to serve as a sensitive biosensoric indicator. In the proposed work, gold nanoparticles were modified with two proteins: i) Borrelia antigen, variable lipoprotein surface-exposed protein (VlsE), and ii) protein A. VlsE antigen induces a strong antibody response against Lyme disease and can be detected from early to late phase during the disease in humans infected with Borrelia. In addition, it shows low cross-reaction with the other non-pathogenic Borrelia strains. The high specificity of VlsE antigen to anti-Borrelia antibodies, combined simultaneously with the high specificity of protein A to the Fc region of all IgG human antibodies, was utilized to develop a rapid test for serological point of care diagnosis of borreliosis in human serum. Only in the presence of anti-Borrelia antibodies in the serum probe, an aggregation of gold nanoparticles can be observed, which is visible by a concentration-dependent colour shift from red (low IgG) to blue (high IgG). Experiments showed it is clearly possible to distinguish between positive and negative sera samples using a simple suspension of the two-protein modified gold nanoparticles in a very short time (30 minutes). The proposed work showed the potential of using such modified gold nanoparticles generally for serological diagnosis. Improved specificity and reduced assay time can be archived in applying increased salt concentrations combined with decreased pH values (pH 5).

Keywords: gold nanoparticles, gold aggregation, serological diagnosis, protein A, lyme borreliosis

Procedia PDF Downloads 398

Authors: Qi LI, Xu Lin, Nengming Lin

Abstract:

Objective: The aim of this study was to investigate the role of desmocollin 2 (DSC2) protein in the proliferation, migration and drug resistance of lung cancer cells. Method: CCK-8 assays and colony formation assays were used to evaluate the effect of dsc2 regulation on cancer cell viability and colony formation. Transwell assays and wound healing assays were also performed. Cell flow double staining was used to detect the apoptosis rate of cells with DSC2, which was added cisplatin. Western blot assay was used to detect cell cycle, PI3k/Akt and apoptosis-related proteins. Results: Our data showed that dsc2 is upregulated in clinical lung cancer tissues compared with pericarcinomatous tissues, and it is differentially expressed in lung cancer cell lines. The down-regulation of dsc2 in A549 and H358 lung cancer cells significantly suppressed the cell proliferation, metastasis, and motility. In contrast, the opposite effects were observed in overexpression of dsc2 both in H23 and PC9 cell lines. In addition to lung adenocarcinoma cell lines, we also examined its expression in lung squamous cell lines, such as H226. Western blotting showed that dsc2 could reduce the level of phosphorylated Akt (Ser 473) and p-mTOR. Thus, it is speculated that dsc2 up-regulation promotes proliferation and invasiveness through activation of the PI3K/AKT pathway. Also, knockdown of dsc2 in A549 and H226 could significantly decreased in the levels of cyclinB and wee1 protein. Additionally, flow cytometry showed that dsc2 knockdown combined with cisplatin could significantly enhance cell apoptosis rate. Conclusion: These data suggest that dsc2 promotes the proliferation and migration of lung cancer cells in vitro. Also, the results suggested that dsc2 could affect the cell cycle and apoptosis of lung cells. Furthermore, knockdown of dsc2 could sensitize cisplatin in both lung adenocarcinoma and lung squamous cell lines. Thus we suggested that dsc2 can be used as a therapeutic target for lung cancer.

Keywords: desmocollin 2, cisplatin, lung cancer, PI3K/AKT, lung squamous cell

Procedia PDF Downloads 76

Authors: Yaw Opoku-Damoah, Run Zhang, Hang Thu Ta, Zhi Ping Xu

Abstract:

Gas therapy is still at an early stage of research and development. Even though most gasotransmitters have proven their therapeutic potential, their handling, delivery, and controlled release have been extremely challenging. This research work employs a versatile nanosystem that is capable of delivering a gasotransmitter in the form of a photo-responsive carbon monoxide-releasing molecule (CORM) for targeted cancer therapy. The therapeutic action was mediated by upconversion nanoparticles (UCNPs) designed to transfer bio-friendly low energy near-infrared (NIR) light to ultraviolet (UV) light capable of triggering carbon monoxide (CO) from a water-soluble amphiphilic manganese carbonyl complex CORM incorporated into a carefully designed lipid drug delivery system. Herein, gaseous CO that plays a role as a gasotransmitter with cytotoxic and homeostatic properties was investigated to instigate cellular apoptosis. After successfully synthesizing the drug delivery system, the ability of the system to encapsulate and mediate the sustained release of CO after light excitation was demonstrated. CO fluorescence probe (COFP) was successfully employed to determine the in vitro drug release profile upon NIR light irradiation. The uptake of nanoparticles enhanced by folates and its receptor interaction was also studied for cellular uptake purposes. The anticancer potential of the final lipid nanoparticle Lipid/UCNPs/CORM/FA (LUCF) was also determined by cell viability assay. Intracellular CO release and a subsequent therapeutic action involving ROS production, mitochondrial damage, and CO production was also evaluated. In all, this current project aims to use in vitro studies to determine the potency and efficiency of a NIR-mediated CORM prodrug delivery system.

Keywords: carbon monoxide-releasing molecule, upconversion nanoparticles, site-specific delivery, amphiphilic manganese carbonyl complex, prodrug delivery system.

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Authors: Ntokozo Dambuza, Maryna Van De Venter, Trevor Koekemoer

Abstract:

Oxidative stress contributes to the pathogenesis of many diseases and consequently antioxidant therapy has attracted much attention as a potential therapeutic strategy. Regardless of the quantities ingested, antioxidants need to reach the diseased tissues at concentrations sufficient to combat oxidative stress. Bioavailability is thus a defining criterion for the therapeutic efficacy of antioxidants. In addition, therapeutic antioxidants must possess biologically relevant characteristics which can target the specific molecular mechanisms responsible for disease related oxidative stress. While many chemical antioxidant assays are available to quantify antioxidant capacity, they relate poorly to the biological environment and provide no information as to the bioavailability. The present comparative study thus aims to characterise green and fermented rooibos extracts, well recognized for their exceptional antioxidant capacity, in terms of antioxidant bioavailability and efficacy in a disease relevant cellular setting. Chinese green tea antioxidant activity was also evaluated. Chemical antioxidant assays (FRAP, DPPH and ORAC) confirmed the potent antioxidant capacity of both green and fermented rooibos, with green rooibos possessing antioxidant activity superior to that of fermented rooibos and Chinese green tea. Bioavailability was assessed using the PAMPA assay and the results indicate that green and fermented rooibos have a permeation coefficient of 5.7 x 10-6 and 6.9 x 10-6 cm/s, respectively. Chinese green tea permeability coefficient was 8.5 x 10-6 cm/s. These values were comparable to those of rifampicin, which is known to have a high permeability across intestinal epithelium with a permeability coefficient of 5 x 10 -6 cm/s. To assess the antioxidant efficacy in a cellular context, U937 and red blood cells were pre-treated with rooibos and Chinese green tea extracts in the presence of a dye DCFH-DA and then exposed to oxidative stress. Green rooibos exhibited highest activity with an IC50 value of 29 μg/ml and 70 μg/ml, when U937 and red blood cells were exposed oxidative stress, respectively. Fermented rooibos and Chinese green tea had IC50 values of 61 μg/ml and 57 μg/ml for U937, respectively, and 221 μg/ml and 405 μg/ml for red blood cells, respectively. These results indicate that fermented and green rooibos extracts were able to permeate the U937 cells and red blood cell membrane and inhibited oxidation of DCFH-DA to a fluorescent DCF within the cells.

Keywords: rooibos, antioxidants, permeability, bioavailability

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Authors: Thomas Louis, Irina Primac, Florent Morfoisse, Tania Durre, Silvia Blacher, Agnes Noel

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In vitro and in vivo lymphangiogenesis assays are essential for the identification of potential lymphangiogenic agents and the screening of pharmacological inhibitors. In the present study, we analyse three biological models: in vitro lymphatic endothelial cell spheroids, in vivo ear sponge assay, and in vivo lymph node colonisation by tumour cells. These assays provide suitable 3D models to test pro- and anti-lymphangiogenic factors or drugs. 3D images were acquired by confocal laser scanning and light sheet fluorescence microscopy. Virtual scan microscopy followed by 3D reconstruction by image aligning methods was also used to obtain 3D images of whole large sponge and ganglion samples. 3D reconstruction, image segmentation, skeletonisation, and other image processing algorithms are described. Fixed and time-lapse imaging techniques are used to analyse lymphatic endothelial cell spheroids behaviour. The study of cell spatial distribution in spheroid models enables to detect interactions between cells and to identify invasion hierarchy and guidance patterns. Global measurements such as volume, length, and density of lymphatic vessels are measured in both in vivo models. Branching density and tortuosity evaluation are also proposed to determine structure complexity. Those properties combined with vessel spatial distribution are evaluated in order to determine lymphangiogenesis extent. Lymphatic endothelial cell invasion and lymphangiogenesis were evaluated under various experimental conditions. The comparison of these conditions enables to identify lymphangiogenic agents and to better comprehend their roles in the lymphangiogenesis process. The proposed methodology is validated by its application on the three presented models.

Keywords: 3D image segmentation, 3D image skeletonisation, cell invasion, confocal microscopy, ear sponges, light sheet microscopy, lymph nodes, lymphangiogenesis, spheroids

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Authors: Hj Mohd Sidek bin Hj Mohd Yusof

Abstract:

More than 1.5 billion people around the world are adversely affected by floods. Floods account for about a third of all natural catastrophes, cause more than half of all fatalities and are responsible for a third of overall economic loss around the world. Giving advanced warning of impending disasters can reduce or even avoid the number of deaths, social and economic hardships that are so commonly reported after the event. Integrated catchment management recognizes that it is not practical or viable to provide structural measures that will keep floodwater away from the community and their property. Non-structural measures are therefore required to assist the community to cope when flooding occurs which exceeds the capacity of the structural measures. Non-structural measures may need to be used to influence the way land is used or buildings are constructed, or they may be used to improve the community’s preparedness and response to flooding. The development and implementation of non-structural measures may be guided and encouraged by policy and legislation, or through voluntary action by the community based on knowledge gained from public education programs. There is a range of non-structural measures that can be used for flood hazard mitigation which can be the use measures includes policies and rules applied by government to regulate the kinds of activities that are carried out in various flood-prone areas, including minimum floor levels and the type of development approved. Voluntary actions taken by the authorities and by the community living and working on the flood plain to lessen flooding effects on themselves and their properties including monitoring land use changes, monitoring and investigating the effects of bush / forest clearing in the catchment and providing relevant flood related information to the community. Response modification measures may include: flood warning system, flood education, community awareness and readiness, evacuation arrangements and recovery plan. A Civil Defense Emergency Management needs to be established for Brunei Darussalam in order to plan, co-ordinate and undertake flood emergency management. This responsibility may be taken by the Ministry of Home Affairs, Brunei Darussalam who is already responsible for Fire Fighting and Rescue services. Several pieces of legislation and planning instruments are in place to assist flood management, particularly: flood warning system, flood education Community awareness and readiness, evacuation arrangements and recovery plan.

Keywords: RTB, radio television brunei, DDMC, district disaster management center, FIR, flood incidence report, PWD, public works department

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Authors: Samiyah Tasleem, Muhammad Abdul Haq, Syed Baqir Shyum Naqvi, Muhammad Abid Husnain, Sajjad Haider Naqvi

Abstract:

The objective of the present study was: a) to investigate the antimicrobial activity of honey samples of different floral sources of Pakistan, b) to recover the phenolic acids in them as a possible contributing factor of antimicrobial activity. Six honey samples from different floral sources, namely: Trachysperm copticum, Acacia species, Helianthus annuus, Carissa opaca, Zizyphus and Magnifera indica were used. The antimicrobial activity was investigated by the disc diffusion method against eight freshly isolated clinical isolates (Staphylococci aureus, Staphylococci epidermidis, Streptococcus faecalis, Pseudomonas aeruginosa, Klebsiella pneumonia, Escherichia coli, Proteus vulgaris and Candida albicans). Antimicrobial activity of honey was compared with five commercial antibiotics, namely: doxycycline (DO-30ug/mL), oxytetracycline (OT-30ug/mL), clarithromycin (CLR–15ug/mL), moxifloxacin (MXF-5ug/mL) and nystatin (NT – 100 UT). The fractions responsible for antimicrobial activity were extracted using ethyl acetate. Solid phase extraction (SPE) was used to recover the phenolic acids of honey samples. Identification was carried out via High-Performance Liquid Chromatography (HPLC). The results indicated that antimicrobial activity was present in all honey samples and found comparable to the antibiotics used in the study. In the microbiological assay, the ethyl acetate honey extract was found to exhibit a very promising antimicrobial activity against all the microorganisms tested, indicating the existence of phenolic compounds. Six phenolic acids, namely: gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids were identified besides some unknown substance by HPLC. In conclusion, Pakistani honey samples showed a broad spectrum antibacterial and promising antifungal activity. Identification of six different phenolic acids showed that Pakistani honey samples are rich sources of phenolic compounds that could be the contributing factor of antimicrobial activity.

Keywords: Pakistani honey, antimicrobial activity, Phenolic acids eg.gallic, caffeic, ferulic, vanillic, benzoic and cinnamic acids

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Authors: Yuda Purwana Roswanjaya, Wouter Kohlen, Rene Geurts

Abstract:

The Trema genus represents a group of fast-growing tropical tree species within the Cannabaceae. Interestingly, five species nested in this lineage -known as Parasponia- can establish rhizobium nitrogen-fixing root nodules, similar to those found in legumes. Parasponia and legumes use a conserved genetic network to control root nodule formation, among which are genes also essential for mycorrhizal symbiosis (the so-called common symbiotic pathway). However, Trema species lost several genes that function exclusively in nodulation, suggesting a loss-of the nodulation trait in Trema. Strikingly, in a Trema orientalis population found in Malaysian Borneo we identified a truncated SYMBIOSIS RECEPTOR KINASE (SYMRK) mutant allele lacking a large portion of the c-terminal kinase domain. In legumes this gene is essential for nodulation and mycorrhization. This raises the question whether Trema orientalis can still be mycorrhized. To answer this question, we established quantitative mycorrhization assay for Parasponia andersonii and Trema orientalis. Plants were grown in closed pots on half strength Hoagland medium containing 20 µM potassium phosphate in sterilized sand and inoculated with 125 spores of Rhizopagus irregularis (Agronutrion-DAOM197198). Mycorrhization efficiency was determined by analyzing the frequency of mycorrhiza (%F), the intensity of the mycorrhizal colonization (%M) and the arbuscule abundance (%A) in the root system. Trema orientalis RG33 can be mycorrhized, though with lower efficiency compared to Parasponia andersonii. From this we conclude that a functional SYMRK kinase domain is not essential for Trema orientalis mycorrhization. In ongoing experiments, we aim to investigate the role of SYMRK in Parasponia andersonii mycorrhization and nodulation. For this two Parasponia andersonii symrk CRISPR-Cas9 mutant alleles were created. One mimicking the TorSYMRKRG33 allele by deletion of exon 13-15, and a full Parasponia andersonii SYMRK knockout.

Keywords: endomycorrhization, Parasponia andersonii, symbiosis receptor kinase (SYMRK), Trema orientalis

Procedia PDF Downloads 164