Search results for: fire assay

Authors: Nahid Eskandari, Marjan Ghagozolo, Ehsan Almasi

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Introduction: Silymarin, as a polyphenolic flavonoid derived from milk thistle (Silybum marianum), is known to have antioxidant, immunomodulatory, antiproliferative, antifibrotic, and antiviral effects. The goal of this study was to determine immunosuppressive effect of Silymarin on proliferation and apoptosis of human T cells in comparison with Rapamycin and FK506. Methods: Peripheral Blood Mononuclear Cells (PBMCs) from healthy individuals were activated with Con A (5µg/ml) and then treated with Silymarin, Rapamycin and FK506 in various concentrations (0.001, 0.01, 0.1, 1, 10,100 and 200M) for 5 days. PBMCs were examined for proliferation using CFSE assay and the concentration that inhibited 50% of the cell proliferation (IC50) was determined for each treatment. For apoptosis assay using flow cytometry, PBMCs were activated with Con A and treated with IC50 dose of Silymarin, Rapamycin and FK506 for 5 days, then cell apoptosis was analysed by FITC-annexin V/PI staining and flow cytometry. The effects of Silymarin, Rapamycin and FK506 on the activation of PARP (poly ADP ribose polymerase) pathway in PBMCs stimulated with Con A and treated with IC50 dose of drugs for 5 days evaluated using the PathScan cleaved PARP sandwich ELISA kit. Results: This study showed that Silymarin had the ability to inhibit T cell proliferation in vitro. Moreover, our results indicated that 100 μM (P < 0.001) and 200 μM (P < 0.001) of Silymarin has more inhibitory effect on T cells proliferation than FK506 and Rapamycin. Our data showed that the effective doses (IC50) of Silymarin, FK506 and Rapamycin were 3×10-5 µM, 10-8 µM and 10-6 µM respectively. Data showed that the inhibitory effect of Silymarin, FK506 and Rapamycin on T cell proliferation was not due to cytotoxicity and none of these drugs at IC50 concentration had not affected the level of cleaved PARP. Conclusion: Silymarin could be a good candidate for immunosuppressive therapy for certain medical conditions with superior efficacy and lesser toxicity in comparison with other immunosuppressive drugs.

Keywords: silymarin, immunosuppressive effect, rapamycin, immunology

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Authors: Racim Rifaat Ferdjani, Zineddine Chetoui

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The goal of our work is to provide a tool that helps control and ensures a global view of the means of firefighting (MLCI) in a food production plant (for example Hamoud Boualem plant). We divided the site into 4 zones, then we identified the firefighting means (MLCI) present in each zone, taking into account their type, weight, location, and fire class as well as their compliance with respect to the regulations in force while assigning them an alphanumeric reference which makes it possible to deduce everything. Thus, the use of a tool in the form of an Excel table was made concrete, and an average compliance rate of 45% was therefore obtained.

Keywords: MLCI, firefighting means, Hamoud, Boualem

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Authors: S. Beekrum, B. Odhav, R. Lalloo, E. A. Amonsou

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Marine microalgae are rich sources of novel and biologically active metabolites; therefore they may be used in the food industry as natural food ingredients and functional foods. They have several biological applications related to health benefits, among others. The aim of the study focused on the screening of phytochemicals from Amphora sp. biomass extracts, and to examine the in vitro antioxidant and antimicrobial potential. Amphora sp. biomass was obtained from CSIR (South Africa) and methanol, hexane and water extracts were prepared. The in vitro antimicrobial effect of extracts were tested against some pathogens (Staphylococcus aureus, Listeria monocytogenes, Bacillus subtilis, Salmonella enteritidis, Escherichia coli, Pseudomonas aeruginosa and Candida albicans), using the disc diffusion assay. Qualitative analyses of phytochemicals were conducted by chemical tests. The present investigation revealed that all extracts showed relatively strong antibacterial activity against most of the tested bacteria. The highest phenolic content was found in the methanolic extract. Results of the DPPH assay showed that the biomass contained strong antioxidant capacity, 79% in the methanolic extract and 85% in the hexane extract. Extracts have displayed effectively reducing power and superoxide anion radical scavenging activity. Results of this study have highlighted potential antioxidant activity in the methanol and hexane extracts. The results of the phytochemical screening showed the presence of terpenoids and sterols with potential applications as food flavorants and functional foods, respectively. The use of Amphora sp. as a natural antioxidant source and a potential source of antibacterial compounds and phytochemicals in the food industry appears promising and should be investigated further.

Keywords: antioxidants, antimicrobial, microalgae, phytochemicals, cymbella

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Authors: Belgheis Ebrahimi, Jun Lu

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In recent years, there has been a growing recognition of the potential of marine-based functional foods and combination therapies in promoting a healthy lifestyle and exploring their effectiveness in preventing or treating diseases. The combination of marine bioactive compounds or extracts offers synergistic or enhancement effects through various mechanisms, including multi-target actions, improved bioavailability, enhanced bioactivity, and mitigation of potential adverse effects. Both the green-lipped mussel (GLM) and fucoidan derived from brown seaweed are rich in bioactivities. These two, mussel and fucoidan, have not been previously formulated together. This study aims to combine GLM oil from Perna canaliculus with low molecular weight fucoidan (LMWF) extracted from Undaria pinnatifida to investigate the unique mixture’s anti-inflammatory and antioxidant properties. The cytotoxicity of individual compounds and combinations was assessed using the MTT assay in (THP-1 and RAW264.7) cell lines. The anti-inflammatory activity of mussel-fucoidan was evaluated by treating LPS-stimulated human monocyte and macrophage (THP1-1) cells. Subsequently, the inflammatory cytokines released into the supernatant of these cell lines were quantified via ELISA. Antioxidant activity was determined by using the free radical scavenging assay (DPPH). DPPH assay demonstrated that the radical scavenging activity of the combinations, particularly at concentrations exceeding 1 mg/ml, showed a significantly higher percentage of inhibition when compared to the individual component. This suggests an enhancement effect when the two compounds are combined, leading to increased antioxidant activity. In terms of immunomodulatory activity, the individual compounds exhibited distinct behaviors. GLM oil displayed a higher ability to suppress the cytokine TNF- compared to LMWF. Interestingly, the LMWF fraction, when used individually, did not demonstrate TNF- suppression. However, when combined with GLM, the TNF- suppression (anti-inflammatory) activity of the combination was better than GLM or LWMF alone. This observation underscores the potential for enhancement interactions between the two components in terms of anti-inflammatory properties. This study revealed that each individual compound, LMWF, and GLM, possesses unique and notable bioactivity. The combination of these two individual compounds results in an enhancement effect, where the bioactivity of each is enhanced, creating a superior combination. This suggests that the combination of LMWF and GLM has the potential to offer a more potent and multifaceted therapeutic effect, particularly in the context of antioxidant and anti-inflammatory activities. These findings hold promise for the development of novel therapeutic interventions or supplements that harness the enhancement effects.

Keywords: combination, enhancement effect, perna canaliculus, undaria pinnatifida

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Authors: Hyun-jin Kim, Gumgyung Gu, Dae-Hyun Ko, Woochang Lee, Sail Chun, Won-Ki Min

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Background: Ovarian carcinoma is the fourth most common cause of cancer-related death in women worldwide. HE4, a novel marker for ovarian cancer could be used for monitoring recurrence or progression of disease in patients with invasive epithelial ovarian carcinoma. It is further intended to be used in conjunction with CA 125 to estimate the risk of epithelial ovarian cancer in women presenting with an adnexal mass. In this study, we aim to evaluate the analytical performance and clinical utility of HE4 assay using Architect i 2000SR(Abbott Diagnostics, USA). Methods: The precision was evaluated according to Clinical and Laboratory Standards Institute(CLSI) EP5 guideline. Three levels of control materials were analyzed twice a day in duplicate manner over 20 days. We calculated within run and total coefficient of variation (CV) at each level of control materials. The linearity was evaluated based on CLSI EP6 guideline. Five levels of calibrator were prepared by mixing high and low level of calibrators. For 43 women with adnexal masses, HE4 and CA 125 were measured and Risk of ovarian malignancy (ROMA) scores were calculated. The patients’ medical records were reviewed to determine the clinical utility of HE4 and ROMA score. Results: In a precision study, the within-run and total CV were 2.0 % and 2.3% for low level of control material, 1.9% and 2.4% for medium level and 0.5 % and 1.1% for high level, respectively. The linear range of HE4 was 14.63 to 1475.15pmol/L. Of the 43 patients, two patients in pre-menopausal group showed the ROMA score above the cut-off level (7.3%). One of them showed CA 125 level within the reference range, while the HE4 was higher than the cut-off. Conclusion: The overall analytical performance of HE4 assay using Architect showed high precision and good linearity within clinically important range. HE4 could be an useful marker for managing patients with adnexal masses.

Keywords: HE4, CA125, ROMA, evaluation, performance

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Authors: Judy M. Obliosca, Olivia Vest, Sandra Poulos, Kelsi Smith, Tammy Ferguson, Abigail Powers Lott, Alicia K. Smith, Yang Xu, Christopher K. Tison

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Post-Traumatic Stress Disorder (PTSD) is a mental health problem that people may develop after experiencing traumatic events such as combat, natural disasters, and major emotional challenges. Tragically, the number of military personnel with PTSD correlates directly with the number of veterans who attempt suicide, with the highest rate in the Army. Research has shown epigenetic risks in those who are prone to several psychiatric dysfunctions, particularly PTSD. Once initiated in response to trauma, epigenetic alterations in particular, the DNA methylation in the form of 5-methylcytosine (5mC) alters chromatin structure and represses gene expression. Current methods to detect DNA methylation, such as bisulfite-based genomic sequencing techniques, are laborious and have massive analysis workflow while still having high error rates. A faster and simpler detection method of high sensitivity and precision would be useful in a clinical setting to confirm potential PTSD etiologies, prevent other psychiatric disorders, and improve military health. A nano-enhanced Surface Plasmon Resonance imaging (SPRi)-based assay that simultaneously detects site-specific 5mC base (termed as PTSD base) in methylated genes related to PTSD is being developed. The arrays on a sensing chip were first constructed for parallel detection of PTSD bases using synthetic and genomic DNA (gDNA) samples. For the gDNA sample extracted from the whole blood of a PTSD patient, the sample was first digested using specific restriction enzymes, and fragments were denatured to obtain single-stranded methylated target genes (ssDNA). The resulting mixture of ssDNA was then injected into the assay platform, where targets were captured by specific DNA aptamer probes previously immobilized on the surface of a sensing chip. The PTSD bases in targets were detected by anti-5-methylcytosine antibody (anti-5mC), and the resulting signals were then enhanced by the universal nanoenhancer. Preliminary results showed successful detection of a PTSD base in a gDNA sample. Brighter spot images and higher delta values (control-subtracted reflectivity signal) relative to those of the control were observed. We also implemented the in-house surface activation system for detection and developed SPRi disposable chips. Multiplexed PTSD base detection of target methylated genes in blood DNA from PTSD patients of severity conditions (asymptomatic and severe) was conducted. This diagnostic capability being developed is a platform technology, and upon successful implementation for PTSD, it could be reconfigured for the study of a wide variety of neurological disorders such as traumatic brain injury, Alzheimer’s disease, schizophrenia, and Huntington's disease and can be extended to the analyses of other sample matrices such as urine and saliva.

Keywords: epigenetic assay, DNA methylation, PTSD, whole blood, multiplexing

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Authors: E. Ormeno, C. Gutigny, J. Ruffault, J. Madrigal, M. Guijarro, C. Lecareux, C. Ballini

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Mediterranean plant species feature numerous metabolic and morpho-physiological responses crucial to survive under both, typical Mediterranean drought conditions and future aggravated drought expected by climate change. Whether these adaptive responses will, in turn, increase the ecosystem perturbation in terms of fire hazard, is an issue that needs to be addressed. The aim of this study was to test whether recurrent and aggravated drought in the Mediterranean area favors the accumulation of waxes in leaf litter, with an eventual increase of litter flammability. The study was conducted in 2017 in a garrigue in Southern France dominated by Quercus coccifera, where two drought treatments were used: a treatment with recurrent aggravated drought consisting of ten rain exclusion structures which withdraw part of the annual precipitation since January 2012, and a natural drought treatment where Q. coccifera stands are free of such structures and thus grow under natural precipitation. Waxes were extracted with organic solvent and analyzed by GC-MS and litter flammability was assessed through measurements of the ignition delay, flame residence time and flame intensity (flame height) using an epiradiator as well as the heat of combustion using an oxygen bomb calorimeter. Results show that after 5 years of rain restriction, wax content in the cuticle of leaf litter increases significantly compared to shrubs growing under natural precipitation, in accordance with the theoretical knowledge which expects increases of cuticle waxes in green leaves in order to limit water evapotranspiration. Wax concentrations were also linearly and positively correlated to litter flammability, a correlation that lies on the high flammability own to the long-chain alkanes (C25-C31) found in leaf litter waxes. This innovative investigation shows that climate change is likely to favor ecosystem fire hazard through accumulation of highly flammable waxes in litter. It also adds valuable information about the types of metabolites that are associated with increasing litter flammability, since so far, within the leaf metabolic profile, only terpene-like compounds had been related to plant flammability.

Keywords: cuticular waxes, drought, flammability, litter

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Authors: Patricia M. B. Saint-Vincent, Anna Furches, Stephanie Galanie, Erica Teixeira Prates, Piet Jones, Nancy Engle, David Kainer, Wellington Muchero, Daniel Jacobson, Timothy J. Tschaplinski

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Uridine diphosphate-glycosyltransferases (UGTs) are enzymes that catalyze sugar transfer to a variety of plant metabolites. UGT substrates, which include plant secondary metabolites involved in lignification, demonstrate new activities and incorporation when glycosylated. Knowledge of UGT function, substrate specificity, and enzyme products is important for plant engineering efforts, especially related to increasing plant biomass through lignification. UGTs in Populus trichocarpa, a biofuel feedstock, and model woody plant, were selected from a pool of gene candidates using rapid prioritization strategies. A functional genomics workflow, consisting of a metabolite genome-wide association study (mGWAS), expression of synthetic codon-optimized genes, and high-throughput biochemical assays with mass spectrometry-based analysis, was developed for determining the substrates and products of previously-uncharacterized enzymes. A total of 40 UGTs from P. trichocarpa were profiled, and the biochemical assay results were compared to predicted mGWAS connections. Assay results confirmed seven of 11 leaf mGWAS associations and demonstrated varying levels of substrate specificity among candidate UGTs. P. trichocarpa UGT substrate processing confirms the role of these newly-characterized enzymes in lignan, flavonoid, and phytohormone metabolism, with potential implications for cell wall biosynthesis, nitrogen uptake, and biotic and abiotic stress responses.

Keywords: Populus, metabolite-gene associations, GWAS, bio feedstocks, glycosyltransferase

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Authors: Enjie Xu, Zhen Huang, Kunpeng Zhu, Jianping Hu, Xiaolong Ma, Yongjie Wang, Jiazhuang Zhu, Chunlin Zhang

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Abstract: Hypoxia and dedifferentiation of osteosarcoma (OS) cells leads to poor prognosis. We plan to identify the role of hypoxia on dedifferentiation and the associated signaling pathways. We performed a sphere formation assay and determined spheroid cells as dedifferentiated cells by detecting stem cell-like markers. RNAi assay was used to explore the expression relationship between hypoxia inducible factor 1 subunit alpha (HIF1A) and platelet derived growth factor receptor beta (PDGFRB). We obtained PDGFRB knockdown and overexpression cells through lentiviral infection experiments and the effects of PDGFRB on cytoskeleton rearrangement and cell adhesion were explored by immunocytochemistry. Wound-healing experiments, transwell assays, and animal trials were employed to investigate the effect of PDGFRB on OS metastasis. Dedifferentiated OS cells were found to exhibit high expression of HIF1A and PDGFRB, and HIF1A promoted the expression of PDGFRB, subsequently activated ras homolog family member A (RhoA), and increased the phosphorylation of myosin light chain (MLC). PDGFRB also enhanced the phosphorylation of focal adhesion kinase (FAK). The OS cell morphology and vinculin distribution were altered by PDGFRB. PDGFRB also promoted cell dedifferentiation and had a significant impact on the metastasis of OS cells both in vitro and in vivo. Our results demonstrated that HIF1A up-regulated PDGFRB under hypoxic conditions, and PDGFRB regulated the actin cytoskeleton by activating RhoA and subsequently phosphorylating MLC, thereby promoting OS dedifferentiation and pulmonary metastasis.

Keywords: osteosarcoma, dedifferentiation, metastasis, cytoskeleton rearrangement, PDGFRB, hypoxia

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Authors: Maryline Vere, Wilma ten Ham-Baloyi, Lucy Ochola, Opeoluwa Oyedele, Lindsey Beyleveld, Siphokazi Tili, Takafira Mduluza, Paula E. Melariri

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Schistosomiasis, caused by Schistosoma (S.) haematobium and Schistosoma (S.) mansoni parasites poses a significant public health challenge in low-income regions. Diagnosis typically relies on identifying specific urine biomarkers such as haematuria, protein, and leukocytes for S. haematobium, while the Point-of-Care Circulating Cathodic Antigen (POC-CCA) assay is employed for detecting S. mansoni. Urinalysis and the POC-CCA assay are favoured for their rapid, non-invasive nature and cost-effectiveness. However, traditional diagnostic methods such as Kato-Katz and urine filtration lack sensitivity in low-transmission areas, which can lead to underreporting of cases and hinder effective disease control efforts. Therefore, in this study, urinalysis and the POC-CCA assay was utilised to diagnose schistosomiasis effectively among school-going children in Nelson Mandela Bay Municipality. This was a cross-sectional study with a total of 759 children, aged 5 to 14 years, who provided urine samples. Urinalysis was performed using urinary dipstick tests, which measure multiple parameters, including haematuria, protein, leukocytes, bilirubin, urobilinogen, ketones, pH, specific gravity and other biomarkers. Urinalysis was performed by dipping the strip into the urine sample and observing colour changes on specific reagent pads. The POC-CCA test was conducted by applying a drop of urine onto a cassette containing CCA-specific antibodies, and the presence of a visible test line indicated a positive result for S. mansoni infection. Descriptive statistics were used to summarize urine parameters, and Pearson correlation coefficients (r) were calculated to analyze associations among urine parameters using R software (version 4.3.1). Among the 759 children, the prevalence of S. haematobium using haematuria as a diagnostic marker was 33.6%. Additionally, leukocytes were detected in 21.3% of the samples, and protein was present in 15%. The prevalence of positive POC-CCA test results for S. mansoni was 3.7%. Urine parameters exhibited low to moderate associations, suggesting complex interrelationships. For instance, specific gravity and pH showed a negative correlation (r = -0.37), indicating that higher specific gravity was associated with lower pH. Weak correlations were observed between haematuria and pH (r = -0.10), bilirubin and ketones (r = 0.14), protein and bilirubin (r = 0.13), and urobilinogen and pH (r = 0.12). A mild positive correlation was found between leukocytes and blood (r = 0.23), reflecting some association between these inflammation markers. In conclusion, the study identified a significant prevalence of schistosomiasis among school-going children in Nelson Mandela Bay Municipality, with S. haematobium detected through haematuria and S. mansoni identified using the POC-CCA assay. The detection of leukocytes and protein in urine samples serves as critical biomarkers for schistosomiasis infections, reinforcing the presence of schistosomiasis in the study area when considered alongside haematuria. These urine parameters are indicative of inflammatory responses associated with schistosomiasis, underscoring the necessity for effective diagnostic methodologies. Such findings highlight the importance of comprehensive diagnostic assessments to accurately identify and monitor schistosomiasis prevalence and its associated health impacts. The significant burden of schistosomiasis in this population highlights the urgent need to develop targeted control interventions to effectively reduce its prevalence in the study area.

Keywords: schistosomiasis, urinalysis, haematuria, POC-CCA

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Authors: Nendouvhada Livhuwani Portia, Nicole Sibuyi, Kwazikwakhe Gabuza, Adewale Fadaka

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Metabolic dysfunction-associated liver disease (MASLD) is a chronic condition characterized by excessive fat accumulation in the liver, distinct from conditions caused by alcohol, viral hepatitis, or medications. MASLD is often linked with metabolic syndrome, including obesity, diabetes, hyperlipidemia, and hypertriglyceridemia. This disease can progress to metabolic dysfunction-associated steatohepatitis (MASH), marked by liver inflammation and scarring, potentially leading to cirrhosis. However, only 43-44% of patients with steatosis develop MASH, and 7-30% of those with MASH progress to cirrhosis. The exact mechanisms underlying MASLD and its progression remain unclear, and there are currently no specific therapeutic strategies for MASLD/MASH. While anti-obesity and anti-diabetic medications can reduce progression, they do not fully treat or reverse the disease. As an alternative, green-synthesized metal nanoparticles (MNPs) are emerging as potential treatments for liver diseases due to their anti-diabetic, anti-inflammatory, and anti-obesity properties with minimal side effects. MNPs like gold nanoparticles (AuNPs) and silver nanoparticles (AgNPs) have been shown to improve metabolic processes by lowering blood glucose, body fat, and inflammation. The study aimed to explore the effects of green-synthesized MNPs on gene expression in an in vitro model of MASH using C3A/HepG2 liver cells. The MASH model was created by exposing these cells to free fatty acids (FFAs) followed by lipopolysaccharide (LPS) to induce inflammation. Cell viability was assessed with the Water-Soluble Tetrazolium (WST)-1 assay, and lipid accumulation was measured using the Oil Red O (ORO) assay. Additionally, mitochondrial membrane potential was assessed by the tetramethyl rhodamine, methyl ester (TMRE) assay, and inflammation was measured with an Enzyme-Linked Immunosorbent Assay (ELISA). The study synthesized AuNPs from Carpobrotus edulis fruit (CeF) and avocado seed (AvoSE) and AgNPs from Salvia africana-lutea (SAL) using optimized conditions. The MNPs were characterized by UV-Vis spectrophotometry and Dynamic Light Scattering (DLS). The nanoparticles were tested at various concentrations for their impact on the C3A/HepG2-induced MASH model. Among the MNPs tested, AvoSE-AuNPs showed the most promise. They reduced cell proliferation and intracellular lipid content more effectively than CeFE-AuNPs and SAL-AgNPs. Molecular analysis using real-time polymerase chain reaction revealed that AvoSE-AuNPs could potentially reverse MASH effects by reducing the expression of key pro-inflammatory and metabolic genes, including tumor necrosis factor-alpha (TNF-α), Fas cell surface death receptor (FAS), Peroxisome proliferator-activated receptor (PPAR)-α, PPAR-γ, and Sterol regulatory element-binding protein (SREBPF)-1. Further research is needed to confirm the molecular mechanisms behind the effects of these MNPs and to identify the specific phytochemicals responsible for their synthesis and bioactivities.

Keywords: gold nanoparticles, green nanotechnology, metal nanoparticles, obesity

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Authors: D. Motlhanka, M. Mine, T. Bagaketse, T. Ngakane

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There is a plethora of evidence from numerous sources that highlights the triumph of naturally derived medicinal plant metabolites with antioxidant capability for repurposed therapeutics. As post-COVID-19 syndromes and non-communicable diseases are on the rise, there is an urgent need to come up with new therapeutic strategies to address the problem. Non-communicable diseases and Post COVID-19 syndromes are classified as socio-economic diseases and are ranked high among threats to health security due to the economic burden they pose to any government budget commitment. Research has shown a strong link between accumulation of free radicals and oxidative stress critical for pathogenesis of non-communicable diseases and COVID-19 syndromes. Botswana has embarked on a robust programme derived from ethno-pharmacognosy and drug repurposing to address these threats to health security. In the current approach, a number of medicinally active plant-derived polyphenolics are repurposed and combined into new medicinal tools to target diabetes, Hypertension, Prostate Cancer and oxidative stress induced Post COVID 19 syndromes such as “brain fog”. All four formulants demonstrated Free Radical scavenging capacities above 95% at 200µg/ml using the diphenylpicryalhydrazyl free radical scavenging assay and the total phenolic contents between 6899-15000GAE(g/L) using the folin-ciocalteau assay respectively. These repurposed medicinal tools offer new hope and potential in the fight against emerging health threats driven by hyper-inflammation and free radical-induced oxidative stress.

Keywords: drug repurposed plant polyphenolics, free radical damage, non-communicable diseases, post COVID 19 syndromes

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Authors: Debasish Das, Mainak Mitra, A.Chaudhuri

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For the purpose of producing moisture vapor permeable waterproof cotton fabric to be used for protective apparel against rain, cotton fabric was coated with the blend of natural rubber and chloroprene rubber containing ammonium acetate as the water-soluble salt, employing a calendar coating technique. Rubber formulations also contained filler, homogenizer, and a typical sulphur curing system. Natural rubber and chloroprene blend in the blend ratio of 30: 70, containing 25 parts of sodium acetate per hundred parts of rubber was coated on the fabric. The coated fabric was vulcanized thereafter at 140oC for 3 h. Coated and vulcanized fabric was subsequently dipped in water for 45 min, followed by drying in air. Such set of treatments produced optimum results. Coated, vulcanized, washed and dried cotton fabric showed optimum developments in the property profiles in respect of waterproofness, breathability as revealed by moisture vapor transmission rate, coating adhesion, tensile properties, abrasion resistance, flex endurance and fire retardancy. Incorporation of highly water-soluble ammonium acetate salt in the coating formulation and their subsequent removal from vulcanized coated layer affected by post washing in consequent to dipping in the water-bath produced holes of only a few microns in the coating matrix of the fabric. Such microporous membrane formed on the cotton fabric allowed only transportation of moisture vapor through them, giving a moisture vapor transmission rate of 3734 g/m2/24h, while acting as a barrier for large liquid water droplet resisting 120cm of the water column in the hydrostatic water-head tester, rendering the coated cotton fabric waterproof. Examination of surface morphology of vulcanized coating by scanning electron microscopy supported the mechanism proposed for development of breathable waterproof layer on cotton fabric by the process employed above. Such process provides an easy and cost-effective route for achieving moisture vapor permeable waterproof cotton.

Keywords: moisture vapour permeability, waterproofness, chloroprene, calendar coating, coating adhesion, fire retardancy

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Authors: Danilo F. Rodrigues, Hérida Regina N. Salgado

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Introduction: The emergence of resistant microorganisms to a large number of clinically approved antimicrobials has been increasing, which restrict the options for the treatment of bacterial infections. As a strategy, drugs with high antimicrobial activities are in evidence. Stands out a class of antimicrobial, the cephalosporins, having as fourth generation cefepime (CEF) a semi-synthetic product which has activity against various Gram-positive bacteria (e.g. oxacillin resistant Staphylococcus aureus) and Gram-negative (e.g. Pseudomonas aeruginosa) aerobic. There are few studies in the literature regarding the development of microbiological methodologies for the analysis of this antimicrobial, so researches in this area are highly relevant to optimize the analysis of this drug in the industry and ensure the quality of the marketed product. The development of microbiological methods for the analysis of antimicrobials has gained strength in recent years and has been highlighted in relation to physicochemical methods, especially because they make possible to determine the bioactivity of the drug against a microorganism. In this context, the aim of this work was the development and validation of a microbiological method for quantitative analysis of CEF in powder lyophilized for injectable solution by turbidimetric assay. Method: For performing the method, Staphylococcus aureus ATCC 6538 IAL 2082 was used as the test microorganism and the culture medium chosen was the Casoy broth. The test was performed using temperature control (35.0 °C ± 2.0 °C) and incubated for 4 hours in shaker. The readings of the results were made at a wavelength of 530 nm through a spectrophotometer. The turbidimetric microbiological method was validated by determining the following parameters: linearity, precision (repeatability and intermediate precision), accuracy and robustness, according to ICH guidelines. Results and discussion: Among the parameters evaluated for method validation, the linearity showed results suitable for both statistical analyses as the correlation coefficients (r) that went 0.9990 for CEF reference standard and 0.9997 for CEF sample. The precision presented the following values 1.86% (intraday), 0.84% (interday) and 0.71% (between analyst). The accuracy of the method has been proven through the recovery test where the mean value obtained was 99.92%. The robustness was verified by the parameters changing volume of culture medium, brand of culture medium, incubation time in shaker and wavelength. The potency of CEF present in the samples of lyophilized powder for injectable solution was 102.46%. Conclusion: The turbidimetric microbiological method proposed for quantification of CEF in lyophilized powder for solution for injectable showed being fast, linear, precise, accurate and robust, being in accordance with all the requirements, which can be used in routine analysis of quality control in the pharmaceutical industry as an option for microbiological analysis.

Keywords: cefepime hydrochloride, quality control, turbidimetric assay, validation

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Authors: Mehrzad Mohabbi Yadollahi, Pouneh Abdollahifard, Behzad Mokhtare, Majid Atashafrazeh

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Geopolymers are believed to provide good fire resistance. The effects of elevated temperatures on mechanical and microstructural properties of pumice-based geopolymer were investigated in this study. Pumice based geopolymer was exposed to elevated temperatures of 200, 400, 600, and 800 ºC for 3 hours. The residual strength of these specimens was determined after cooling at room temperature and microstructures of these samples were investigated by FTIR and SEM analyses. Specimens which were initially grey turned reddish accompanied by the appearance of cracks as temperatures increased to 600 and 800 ºC.

Keywords: geopolymer, pumice, elevated temperature, SEM, FTIR

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Authors: Shimayali Kaushal, Nitesh Priyadarshi, Nitin Kumar Singhal

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Food borne bacterial species have been identified as major pathogens in most of the severe pathogen-related diseases among humans which result in great loss to human health and food industry. Conventional methods like plating and enzyme-linked immune sorbent assay (ELISA) are time-consuming, laborious and require specialized instruments. Nanotechnology has emerged as a great field in case of rapid detection of pathogens in recent years. The AuNRs material has good electro-optical properties due to its larger light absorption band and scattering in surface plasmon resonance wavelength regions. By exploiting the sugar-based adhesion properties of microorganism, we can use the glycoconjugates capped gold nanorods as a potential nanobiosensor to detect the foodborne pathogen. In the present study, polyethylene glycol (PEG) coated gold nanorods (AuNRs) were prepared and functionalized with different types of carbohydrates and further characterized by UV-Visible spectrophotometry, dynamic light scattering (DLS), transmission electron microscopy (TEM). The reactivity of above said nano-biosensor was probed by lectin binding assay and also by different strains of foodborne bacteria by using spectrophotometric and microscopic techniques. Due to the specific interaction of probe with foodborne bacteria (Escherichia coli, Pseudomonas aeruginosa), our nanoprobe has shown significant and selective ablation of targeted bacteria. Our findings suggest that our nanoprobe can be an ideal candidate for selective optical detection of food pathogens and can reduce loss to the food industry.

Keywords: glyco-conjugates, gold nanorods, nanobiosensor, nanoprobe

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Authors: Devinder Kaur Sugga, Ekamdeep Kaur, Jaspreet Kaur, C. Rajesh, Preeti Rajesh, Harsimran Kaur

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Withanolides are steroidal lactones and are highly oxygenated phytoconstituents that can be developed as potential anti-carcinogenic agents. The two main withanolides, namely Withaferin A and Withanolides D, have been extensively studied for their pharmacological activities. Both these withanolides are present in the Withania somnifera (WS) leaves belonging to the family Solanaceae, also known as “Indian ginseng .”In this study effects of WS leaf extract on the MCF7 breast cancer cell line were investigated by performing a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to evaluate the cytotoxic effects and in vitro wound-healing assay to study the effect on cancer cell migration. Our data suggest WS extracts have cytotoxic effects and are effective anti-migrating agents and thus can be a source of potential candidates for the development of potential agents against metastasis. Thus, it can be a source of potential candidates for the development of potential agents against metastasis. Insight into these results, the in-silico approach to identify the possible protein targets interacting with withanolides was taken. Protein kinase C alpha (PKCα) was among the selected 5 top-ranked target proteins identified by the Swiss Target Prediction tool. PKCα is known to promote the growth and invasion of cancer cells and is being evaluated as a prognostic biomarker and therapeutic target in clinically aggressive tumors. Molecular docking of Withaferin A and Withanolides D was performed using AutoDock Vina. Both the bioactive compounds interacted with PKCα. The targets predicted using this approach will serve as leads for the possible therapeutic potential of withanolides, the bioactive ingredients of WS extracts, as anti-cancer drugs.

Keywords: withania somnifera, withaferin A, withanolides D, PKCα

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Authors: J. Harrison Kurunathan, R. Jayaparvathy

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"Assistive Robotics" is the field that deals with the study of robots that helps in human motion and also empowers human abilities by interfacing the robotic systems to be manipulated by human motion. The proposed model is a robotic arm that works as a haptic interface on the basis on accelerometers and DC motors that will function with respect to the movement of the human muscle. The proposed model would effectively work as a haptic interface that would reduce human effort in the field of defense and rescue. This can be used in very critical conditions like fire accidents to avoid causalities.

Keywords: accelerometers, haptic interface, servo motors, signal processing

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Authors: Vishnu Varthan Vaithiyalingamjagannathan, M. N. Sathishkumar, K. S. Lakhsmi, D. Satheeshkumar, Srividyaammayappanrajam

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Background:Naringenin, aglycone flavonoid possess certain activities like anti-oxidant, anti-estrogenic, anti-diabetic, cardioprotective, anti-obesity,anti-inflammatory, hepatoprotective and also have anti-cancer characteristics like carcinogenic inactivation, cell cycle arrest, anti-proliferation, apoptosis, anti-angiogenesis and enhances anti-oxidant activity. Methodology:The inhibitory effect of Naringenin in lung tumor progression estimated with adenocarcinoma (A549) cell lines (in vitro) and C57BL/6 mice injected with 5 X 106A549 cell lines (in vivo) in a tri-dose manner (Naringenin 100mg/kg,150mg/kg, and 200mg/kg) compared with standard chemotherapy drug cisplatin (7mg/kg). Results:The results of the present study revealed a dose-dependent activity in Naringenin and combination with cisplatin at a higher dose which showed decreased tumor progression in mice. In vitro studies carried out for estimation of cell survival and Nitric Oxide (NO) level, shows dose dependent action of Naringenin with IC50 value of 42µg/ml. In vivo studies were carried out in C57BL/6 mice. Naringenin satisfied the condition of an anti-cancer molecule with its characteristics in fragmentation assay, Zymography assay, anti-oxidant, and myeloperoxidase studies, than cisplatin which failed in anti-oxidant and myeloperoxidase effect. Both in vitro and in vivo establishes dose dependent decrease in NO levels. But whereas, Naringenin showed adverse results in Matrix Metalloproteinase (MMP) enzymatic levels with increase in dose levels. Conclusion:From the present study, Naringenin could suppress the lung tumor progression when given individually and also in combinatorial with standard chemotherapy drug.

Keywords: naringenin, in vitro, cell line, anticancer

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Authors: Ewa Kicko Walczak, Grazyna Rymarz

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In recent years, nanotechnology has been successfully applied for flame retardancy of polymers, in particular for construction materials. The consumption of thermoset resins as a construction polymers materials is approximately over one million tone word wide. Excellent mechanical, relatively high heat and thermal stability of their type of polymers are proven for variety applications, e.g. transportation, electrical, electronic, building part industry. Above applications in addition to the strength and thermal properties also requires -referring to the legal regulation or recommendation - an adequate level of flammability of the materials. This publication present the evaluation was made of effectiveness of flame retardancy of halogen-free hybrid flame retardants(FR) as compounds nitric/phosphorus modifiers that act with nanofillers (nano carbons, organ modified montmorillonite, nano silica, microsphere) in relation to unsaturated polyester/epoxy resins and glass-reinforced on base this resins laminates(GRP) as a final products. The analysis of the fire properties provided proof of effective flame retardancy of the tested composites by defining oxygen indices values (LOI), with the use of thermogravimetric methods (TGA) and combustion head (CH). An analysis of the combustion process with Cone Calorimeter (CC) method included in the first place N/P units and nanofillers with the observed phenomenon of synergic action of compounds. The fine-plates, phase morphology and rheology of composites were assessed by SEM/ TEM analysis. Polymer-matrix glass reinforced laminates with modified resins meet LOI over 30%, reduced in a decrease by 70% HRR (according to CC analysis), positive description of the curves TGA and values CH; no adverse negative impact on mechanical properties. The main objective of our current project is to contribute to the general understanding of the flame retardants mechanism and to investigate the corresponding structure/properties relationships. We confirm that nanotechnology systems are successfully concept for commercialized forms for non-flammable GRP pipe, concrete composites, and flame retardant tunnels constructions.

Keywords: fire retardants, FR, halogen-free FR nanofillers, non-flammable pipe/concrete, thermoset resins

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Authors: J. Vinay , Kusumbati Besra, Niharika Pattnaik, Shivaram Prasad Singh, Manjusha Dixit

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Gallbladder cancer (GBC) is rare but highly malignant cancer; its prevalence is more in certain geographical regions and ethnic groups, which include the Northern and Eastern states of India. Previous studies in India have reported genetic predisposition as one of the risk factors in GBC pathogenesis. Although the matrix metalloproteinase-14 (MMP14) is a well-known modulator of the tumor microenvironment and tumorigenesis and TCGA data also suggests its upregulation yet, its role in the genetic predisposition for GBC is completely unknown. We elucidated the role of MMP14 promoter variants as genetic risk factors and their implications in expression modulation. We screened MMP14 promoter variants association with GBC using Sanger’s sequencing in approximately 300 GBC and 300 control subjects and 26 GBC tissue samples of Indian ethnicity. The immunohistochemistry was used to check the MMP14 protein expression in GBC tissue samples. The role of promoter variants on expression levels was elucidated using a luciferase reporter assay. The variants rs1004030 (p-value = 0.0001) and rs1003349 (p-value = 0.0008) were significantly associated with gallbladder cancer. The luciferase assay in two different cell lines, HEK-293 (p = 0.0006) and TGBC1TKB (p = 0.0036) showed a significant increase in relative luciferase activity in the presence of risk alleles for both the single nucleotide polymorphisms (SNPs). Similarly, genotype-phenotype correlation in patients samples confirmed that the presence of risk alleles at rs1004030 and rs1003349 increased MMP14 expression. Overall, this study unravels the genetic association of MMP14 promoter variants with gallbladder cancer, which may contribute to pathogenesis by increasing its expression.

Keywords: gallbladder cancer, matrix metalloproteinase-14, single nucleotide polymorphism, case control study, genetic association study

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Authors: Naveen Kumar B. T., Anuj Tyagi, Niraj Kumar Singh, Visanu Boonyawiwat, Shanthanagouda A. H., Orawan Boodde, Shankar K. M., Prakash Patil, Shubhkaramjeet Kaur

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Since 2009, Acute Hepatopancreatic Necrosis Disease (AHPND) outbreaks have increased rapidly, and these have led to the major economic losses to the global shrimp industry. In comparison to other treatments, passive immunity and monoclonal antibody (MAb) based farmer level kit have proved their importance in controlling and treating the diseases in the shrimp industry. In the present study, MAbs were produced against the recombinant PirB protein Vibrio parahaemolyticus strain causing AHPND. Briefly, Balb/C mice were immunized with rPirB at 15 days interval, and antibody titer was determined by ELISA. Spleen cells from mice showing high antibody titer were fused with SP2O myeloma cells for hybridoma production. Among 130 hybridomas, four showed high antibody titer and positive reactivity in an immunoblot assay. In Western blot assay, three out of four MAbs (4C4, 2C2 and 4G3) showed reactivity to rPirB protein. However, in the natural host, only Mab clone 4G3 show strong reactivity (with a strain of V. parahemolyticus causing EMS/AHPND). These clones also showed reactivity with less than 20 kDa proteins in AHPND free V. parahaemolyticus (Thailand stain). Further, on from MAb 4G3 clone, four panels of single cell MAbs clones (G3F5, G3B8, G3H2, and G3D6) were produced of which three showed strong positive reactivity to rPirB protein in the Western blot. These MAbs have potential for controlling and prevention of the AHPND through passive immunity and development of filed level rapid diagnostic kits.

Keywords: shrimp, economic loss, AHPND, MAb

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Authors: Tugba Altin

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In an era where the impacts of climate change resonate more pronouncedly than ever, communities globally grapple with events bearing both tangible and intangible ramifications. Situating this within the evolving landscapes of Psychological and Behavioral Sciences, this research probes the profound psychological and behavioral responses evoked by such events. The Lytton Creek Fire of 2021 epitomizes these challenges. While tangible destruction is immediate and evident, the intangible repercussions—emotional distress, disintegration of cultural landscapes, and disruptions in place attachment (PA)—require meticulous exploration. PA, emblematic of the emotional and cognitive affiliations individuals nurture with their environments, emerges as a cornerstone for comprehending how environmental cataclysms influence cultural identity and bonds to land. This study, harmonizing the core tenets of an interpretive phenomenological approach with a hermeneutic framework, underscores the pivotal nature of this attachment. It delves deep into the realm of individuals' experiences post the Lytton Creek Fire, unraveling the intricate dynamics of PA amidst such calamity. The study's methodology deviates from conventional paradigms. Instead of traditional interview techniques, it employs walking audio sessions and photo elicitation methods, granting participants the agency to immerse, re-experience, and vocalize their sentiments in real-time. Such techniques shed light on spatial narratives post-trauma and capture the otherwise elusive emotional nuances, offering a visually rich representation of place-based experiences. Central to this research is the voice of the affected populace, whose lived experiences and testimonies form the nucleus of the inquiry. As they renegotiate their bonds with transformed environments, their narratives reveal the indispensable role of cultural landscapes in forging place-based identities. Such revelations accentuate the necessity of integrating both tangible and intangible trauma facets into community recovery strategies, ensuring they resonate more profoundly with affected individuals. Bridging the domains of environmental psychology and behavioral sciences, this research accentuates the intertwined nature of tangible restoration with the imperative of emotional and cultural recuperation post-environmental disasters. It advocates for adaptation initiatives that are rooted in the lived realities of the affected, emphasizing a holistic approach that recognizes the profundity of human connections to landscapes. This research advocates the interdisciplinary exchange of ideas and strategies in addressing post-disaster community recovery strategies. It not only enriches the climate change discourse by emphasizing the human facets of disasters but also reiterates the significance of an interdisciplinary approach, encompassing psychological and behavioral nuances, for fostering a comprehensive understanding of climate-induced traumas. Such a perspective is indispensable for shaping more informed, empathetic, and effective adaptation strategies.

Keywords: place attachment, community recovery, disaster response, restorative landscapes, sensory response, visual methodologies

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Authors: Seyede Sanaz Seyedebrahimi, Shima Rahimi, Shohreh Fakhari, Ali Jalili

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Background: CXCR4, as a chemokine receptor, plays well-known roles in various types of cancers. Several studies have been conducted to overcome CXCR4 axis acts in multiple myeloma (MM) pathogenesis and progression. Dexamethasone, a standard treatment for multiple myeloma, has been shown to increase CXCR4 levels in multiple myeloma cell lines. Herein, we focused on the effects of metformin and dexamethasone on CXCR4 at the cellular level and the migration rate of cell lines after exposure to a combination compared to single-agent models. Materials and Method: Multiple myeloma cell lines (U266 and RPMI8226) were cultured with different metformin and dexamethasone concentrations in single-agent and combination models. The simultaneous combination doses were calculated by CompuSyn software. Cell surface and mRNA expression of CXCR4 were determined using flow cytometry and the quantitative reverse transcription-polymerase chain reaction (qRT-PCR) assay, respectively. The Transwell cell migration assay evaluated the migration ability. Results: In concurred with previous studies, our results showed a dexamethasone up-regulation effect on CXCR4 in a dose-dependent manner. Although, the metformin single-agent model could reduce CXCR4 expression of U266 and RPMI8226 in cell surface and mRNA expression level. Moreover, the administration of metformin and dexamethasone simultaneously exerted a higher suppression effect on CXCR4 expression than the metformin single-agent model. The migration rate through the combination model's matrigel membrane was remarkably lower than the metformin and dexamethasone single-agent model. Discussion: According to our findings, the combination of metformin and dexamethasone effectively inhibited dexamethasone-induced CXCR4 expression in multiple myeloma cell lines. As a result, metformin may be counted as an alternative medicine combined with other chemotherapies to combat multiple myeloma. However, more research is required.

Keywords: CXCR4, dexamethasone, metformin, migration, multiple myeloma

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Authors: Deena Ramful-Baboolall, Eshana B. N. Bhatoo

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Morinda citrifolia L., commonly known as noni fruit, is rich in phytochemicals. This study investigated the phytophenolics content and antioxidant activity of green, mature green and ripe noni fruits. The vitamin C content ranged from 41.12 ± 0.083 to 143.63 ± 0.146 mg / 100 ml in fresh noni fruits. Ripe fruits contained the highest level of ascorbic acid followed by mature green and green fruits (p < 0.05). The total phenol content ranged from 0.909 (green) to 2.305 (ripe) mg / g of FW whilst the total flavonoid content ranged from 1.054 (green) to 2.116 (ripe) mg/g of FW. The in vitro antioxidant activity of the Morinda citrifolia L. extracts was also analysed using FRAP and TEAC assays. The reducing power of the fruit extracts as assessed by the FRAP assay decreased in the following order: ripe > mature green > green (p < 0.05). The TEAC values ranged from 0.2631 to 0.8921 µmol / g FW, with extracts of fruits at the mature green stage having highest values followed by fruits at the ripe and green stage respectively (p < 0.05). High correlation values were obtained between total phenolics, total flavonoids, ascorbic acid contents and the TEAC and FRAP assays (r > 0.8). Noni fruit extracts (0.2 and 0.4 % m / m) were compared with BHT (0.02 % m / m) on their ability to protect canola oil and mayonnaise, prepared with canola oil, against lipid oxidation during storage at 40°C. Mature green and ripe extracts, at both concentrations, were more effective than BHT in retarding oxidation in both food systems as evidenced by peroxide value and conjugated diene value determinations. Noni extracts were also very effective in inhibiting lipid peroxidation in tuna fish homogenates, assessed using TBARS assay. Noni fruits at the mature green and ripe stages represent a potential source of natural antioxidants for use a food additive.

Keywords: antioxidant, canola oil, mayonnaise, Morinda citrifolia L. fruit extracts, total flavonoids, total phenol

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Authors: Ken Munro

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Cruise ships are possibly the most complex collection of systems it is possible to find in one physical, moving location. Propulsion, navigation, power generation and more, combined with a hotel, restaurant, casino, theatre etc, with safety and fire control systems to boot. That complexity creates huge challenges with keeping OT and IT systems apart. Ships engines are often remotely managed, network segregation is often defeated through troubleshooting when at sea. This session will refer to multiple entertaining and informative tales of taking control of ships, including accessing a ships Azipods via a game simulator for passengers. Fortunately, genuine attacks against vessels are very rare, but the effects and impacts to world trade are becoming increasingly obvious.

Keywords: maritime security, cybersecurity, OT, IT, networks

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Authors: Faraj M. Elkhatri, Hana Ellafi

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In recent years, there has been a growing recognition of the potential of marine-based functional foods and combination therapies in promoting a healthy lifestyle and exploring their effectiveness in preventing or treating diseases. The combination of marine bioactive compounds or extracts offers synergistic or enhancement effects through various mechanisms, including multi-target actions, improved bioavailability, enhanced bioactivity, and mitigation of potential adverse effects. Both the green-lipped mussel (GLM) and fucoidan derived from brown seaweed are rich in bioactivities. These two, mussel and fucoidan, have not been previously formulated together. This study aims to combine GLM oil from Perna canaliculus with low molecular weight fucoidan (LMWF) extracted from Undaria pinnatifida to investigate the unique mixture’s anti-inflammatory and antioxidant properties. The cytotoxicity of individual compounds and combinations was assessed using the MTT assay in (THP-1 and RAW264.7) cell lines. The anti-inflammatory activity of mussel-fucoidan was evaluated by treating LPS-stimulated human monocyte and macrophage (THP1-1) cells. Subsequently, the inflammatory cytokines released into the supernatant of these cell lines were quantified via ELISA. Antioxidant activity was determined by using the free radical scavenging assay (DPPH). DPPH assay demonstrated that the radical scavenging activity of the combinations, particularly at concentrations exceeding 1 mg/ml, showed a significantly higher percentage of inhibition when compared to the individual component. This suggests an enhancement effect when the two compounds are combined, leading to increased antioxidant activity. In terms of immunomodulatory activity, the individual compounds exhibited distinct behaviors. GLM oil displayed a higher ability to suppress the cytokine TNF- compared to LMWF. Interestingly, the LMWF fraction, when used individually, did not demonstrate TNF- suppression. However, when combined with GLM, the TNF- suppression (anti-inflammatory) activity of the combination was better than GLM or LWMF alone. This observation underscores the potential for enhancement interactions between the two components in terms of anti-inflammatory properties. This study revealed that each individual compound, LMWF, and GLM, possesses unique and notable bioactivity. The combination of these two individual compounds results in an enhancement effect, where the bioactivity of each is enhanced, creating a superior combination. This suggests that the combination of LMWF and GLM has the potential to offer a more potent and multifaceted therapeutic effect, particularly in the context of antioxidant and anti-inflammatory activities. These findings hold promise for the development of novel therapeutic interventions or supplements that harness the enhancement effects.

Keywords: formation damage, porosity loses, pore throat, quartz cement

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Authors: Mudassar Mohiuddin, Zahid Iqbal

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Introduction: Clostridium perfringens is an important pathogen responsible for causing enteric diseases in both human and animals. The bacteria produce several toxins. These toxins play vital role in the pathogenesis of various fatal enteric diseases and are classified into five types, on the basis of the differential production of Alpha, Beta, Epsilon and Iota toxins. In addition to the so-called major toxins, there are other toxins like beta2 toxin, produced by some strains of C. perfringens which may play a role in the pathogenesis of disease. Aim of the study: In this study a multiplex PCR assay was developed and used for detection of cpb2 gene to identify the Beta2 harboring isolates among different types of C. perfringens. Objectives: The primary objective of this study was to identify the prevalence of β2-toxin gene in local isolates of Clostridium perfringens. Methodology: This was an experimental study. Random sampling technique was used. A total of 97 sheep and goats were included in this study. All were Pakistani local breeds. The samples were collected during the period from Sep, 2014 to Mar, 2015 from selected districts of Punjab province (Pakistan). Faecal samples were cultured in cooked meat media. The identification of Clostridium perfringens was made on the basis of biochemical tests. Multiplex PCR was performed to identify the toxin genes. Results: A total of 43 C. perfringens isolates were genotyped using multiplex PCR assay. The gene encoding C. perfringens β2-toxin (cpb2) was present in more than 50% of the isolates genotyped. However, the prevalence of this gene varied between sheep and goat isolates. Conclusion: The present study suggests the high occurrence of C. perfringens b2-toxin (cpb2) in the local isolates of Pakistan. As β2-toxin is present in both healthy and diseased animals, so further studies are suggested to establish the role of β2-toxin in pathogenesis of the clostridial enteric diseases.

Keywords: beta 2 toxin gene, clostridium perfringens, enteric diseases, goats, multiplex PCR, sheep

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Authors: Zeina Bou Diab, Arij Daou

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The neural mechanisms of sequential behaviors are intensively studied, with songbirds a focus for learned vocal production. We are studying the premotor nucleus HVC at a nexus of multiple pathways contributing to song learning and production. The HVC consists of multiple classes of neuronal populations, each has its own cellular, electrophysiological and functional properties. During singing, a large subset of motor cortex analog-projecting HVCRA neurons emit a single 6-10 ms burst of spikes at the same time during each rendition of song, a large subset of basal ganglia-projecting HVCX neurons fire 1 to 4 bursts that are similarly time locked to vocalizations, while HVCINT neurons fire tonically at average high frequency throughout song with prominent modulations whose timing in relation to song remains unresolved. This opens the opportunity to define models relating explicit HVC circuitry to how these neurons work cooperatively to control learning and singing. We developed conductance-based Hodgkin-Huxley models for the three classes of HVC neurons (based on the ion channels previously identified from in vitro recordings) and connected them in several physiologically realistic networks (based on the known synaptic connectivity and specific glutaminergic and gabaergic pharmacology) via different architecture patterning scenarios with the aim to replicate the in vivo firing patterning behaviors. We are able, through these networks, to reproduce the in vivo behavior of each class of HVC neurons, as shown by the experimental recordings. The different network architectures developed highlight different mechanisms that might be contributing to the propagation of sequential neural activity (continuous or punctate) in the HVC and to the distinctive firing patterns that each class exhibits during singing. Examples of such possible mechanisms include: 1) post-inhibitory rebound in HVCX and their population patterns during singing, 2) different subclasses of HVCINT interacting via inhibitory-inhibitory loops, 3) mono-synaptic HVCX to HVCRA excitatory connectivity, and 4) structured many-to-one inhibitory synapses from interneurons to projection neurons, and others. Replication is only a preliminary step that must be followed by model prediction and testing.

Keywords: computational modeling, neural networks, temporal neural sequences, ionic currents, songbird

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Authors: P. Jost, L. Muckova, M. Matula, J. Pejchal, D. Jun, R. Stetina

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Sulfur mustard (bis(2-chlorethyl) sulfide) is highly toxic, chemical warfare agent that has been used in the past in several armed conflicts. Except for the skin, respiratory tract is one of the important routes of exposure. The elucidation and understanding of the mechanism of toxicity of SM have been effort intensive research. The multiple targets character of SM caused cellular damage resulted in activation of many different mechanisms which contribute to cellular response and participate in the final cytopathology effect. In our present work, we compared time-dependent changes in sulfur mustard exposed adult human lung fibroblasts NHLF and lung epithelial alveolar cell line A-549. Cell viability (MTT assay, Calcein-AM assay, and xCELLigence - real-time cell analysis), apoptosis (flow cytometry), mitochondrial membrane potential (Δψm, flow cytometry), reactive oxygen species induction (DC and cell cycle distribution (flow cytometry) were studied. We observed significantly decreased mitochondrial membrane potential and subsequent induction of apoptosis correlating with decreased cellular viability in the sulfur mustard exposed cells. In low concentrations, sulfur mustard-induced S-phase cell cycle arrest, on the other hand, high concentrations, cell cycle phase distribution of sulfur mustard exposed cells resembled cell cycle phase distribution of control group, which implies nonspecific cell cycle inhibition. Epithelial cells A-549 was found as more sensible to sulfur mustard toxicity. Acknowledgements: This work was supported by a long-term organization development plan Medical Aspects of Weapons of Mass Destruction of the Faculty of Military Health Sciences, University of Defence.

Keywords: apoptosis, cell cycle, cytotoxicity, sulfur mustard

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