Extraction and partial purification of Aspergillus flavus cell wall associated saponin hydrolase

In spite of the importance of saponin hydrolase (SH) enzyme, in the production of biologically active compounds from natural saponins, it is surprising that many aspects of its nature are unknown. The results of the present work revealed that Aspergillus flavus was capable of expressing three SH forms; extracellular, intracellular and cell wall-bound forms. SH cell bound enzyme constituted to more than 75% of the total enzymatic activity in the production medium. The sequential extraction process of SH cell bound enzyme revealed that 47.5% of SH was cytosolic and the rest (52.5%) was associated with the cell wall. The highest SH extraction yield was achieved when 0.25 M Tris-HCl lysis buffer supplemented with 1% Triton X-100 for 24 h at 4-25 °C and pH 8 were applied. Under these optimized conditions, A. flavus SH yield increased from 23.6 to 85.83%. The partial purification was achieved by applying successively acetone precipitation, lyophilization, dialysis, and anion exchange chromatography on Fractogel EMD DEAE-650S to the extract. The specific activity of the enzyme extract was 0.27 U/mg after 75% acetone fractionation, while that after anion exchange chromatography was 0.65 U/mg protein. The final enzyme preparation was 7.3-fold purer than the crude extract

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Surgical site infection after gastrointestinal surgery in high-income, middle-income, and low-income countries: a prospective, international, multicentre cohort study

Background: Surgical site infection (SSI) is one of the most common infections associated with health care, but its importance as a global health priority is not fully understood. We quantified the burden of SSI after gastrointestinal surgery in countries in all parts of the world. Methods: This international, prospective, multicentre cohort study included consecutive patients undergoing elective or emergency gastrointestinal resection within 2-week time periods at any health-care facility in any country. Countries with participating centres were stratified into high-income, middle-income, and low-income groups according to the UN's Human Development Index (HDI). Data variables from the GlobalSurg 1 study and other studies that have been found to affect the likelihood of SSI were entered into risk adjustment models. The primary outcome measure was the 30-day SSI incidence (defined by US Centers for Disease Control and Prevention criteria for superficial and deep incisional SSI). Relationships with explanatory variables were examined using Bayesian multilevel logistic regression models. This trial is registered with ClinicalTrials.gov, number NCT02662231. Findings: Between Jan 4, 2016, and July 31, 2016, 13 265 records were submitted for analysis. 12 539 patients from 343 hospitals in 66 countries were included. 7339 (58·5%) patient were from high-HDI countries (193 hospitals in 30 countries), 3918 (31·2%) patients were from middle-HDI countries (82 hospitals in 18 countries), and 1282 (10·2%) patients were from low-HDI countries (68 hospitals in 18 countries). In total, 1538 (12·3%) patients had SSI within 30 days of surgery. The incidence of SSI varied between countries with high (691 [9·4%] of 7339 patients), middle (549 [14·0%] of 3918 patients), and low (298 [23·2%] of 1282) HDI (p < 0·001). The highest SSI incidence in each HDI group was after dirty surgery (102 [17·8%] of 574 patients in high-HDI countries; 74 [31·4%] of 236 patients in middle-HDI countries; 72 [39·8%] of 181 patients in low-HDI countries). Following risk factor adjustment, patients in low-HDI countries were at greatest risk of SSI (adjusted odds ratio 1·60, 95% credible interval 1·05–2·37; p=0·030). 132 (21·6%) of 610 patients with an SSI and a microbiology culture result had an infection that was resistant to the prophylactic antibiotic used. Resistant infections were detected in 49 (16·6%) of 295 patients in high-HDI countries, in 37 (19·8%) of 187 patients in middle-HDI countries, and in 46 (35·9%) of 128 patients in low-HDI countries (p < 0·001). Interpretation: Countries with a low HDI carry a disproportionately greater burden of SSI than countries with a middle or high HDI and might have higher rates of antibiotic resistance. In view of WHO recommendations on SSI prevention that highlight the absence of high-quality interventional research, urgent, pragmatic, randomised trials based in LMICs are needed to assess measures aiming to reduce this preventable complication

Poly-3-Methyl Aniline-Assisted Spherical PbS Quantum Dots through the Ionic Adsorption Deposition Method as a Novel and Highly Efficient Photodetector in UV, Vis, and NIR Regions

This study describes the preparation and characterization of glass/poly-3-methyl aniline (P3MA)/PbS quantum dot (QD) optoelectronic photodetector to detect and sense the light in broad spectral regions of UV, Vis, and NIR. This work is carried out to solve the drawbacks of other studs that prepare detectors in just one or two optical regions. Previous studies have used high-priced techniques. The deposition of P3MA on the glass surface was carried out by in situ oxidation process. Then, this polymer film was used to assist the deposition of PbS-QD particles through the ionic adsorption deposition method. The latter was performed using four different concentrations of Pb(NO3)2 solution (0.01, 0.03, 0.05, and 0.07 M) to form four P3MA/PbS composites: I, II, III, and IV, respectively. The chemical structure, morphologies, and electrical and optical properties of these composites were determined using different analytical tools. The SEM confirmed the formation of spherical QD particles of PbS on the P3MA surface. The TEM analysis showed that the composite has an average size of 5 nm, with the interatomic distances of 0.4 nm. Furthermore, the optical band gap values were 1.53, 1.52, 1.50, and 1.51 eV, respectively. The optoelectronic device could detect and sense light from 390 to 636 nm under various optical wavelengths. The produced current density (Jph) values decreased from 0.029 mA.cm-2 at 390 nm to 0.022 mA.cm-2 at 500 nm and then increased until 0.024 mA.cm-2 at 636 nm. The light sensing was determined through the photoresponsivity (R) and detectivity (D) parameters, in which the photodetector has R and D values of 0.29 mA.cm-2 and 6.5×107 Jones, respectively. Finally, a simple mechanism was proposed to explain the light sensing through the prepared optoelectronic device. Soon, our team works on the industrial applications of this optoelectronic device in the industry field related to the great optoelectronic device technical properties and its low cost and easy preparation

Characterization, modeling, and anticancer activity of L.arginase production from marine Bacillus licheniformis OF2

Abstract Background L-arginase, is a powerful anticancer that hydrolyzes L-arginine to L-ornithine and urea. This enzyme is widely distributed and expressed in organisms like plants, fungi, however very scarce from bacteria. Our study is based on isolating, purifying, and screening the marine bacteria that can produce arginase. Results The highest arginase producing bacteria will be identified by using microbiological and molecular biology methods as Bacillus licheniformis OF2. Characterization of arginase is the objective of this study. The activity of enzyme was screened, and estimated beside partial sequencing of arginase gene was analyzed. In silico homology modeling was applied to generate the protein's 3D structure, and COACH and COFACTOR were applied to determine the protein's binding sites and biological annotations based on the I-TASSER structure prediction. The purified enzyme was undergone an in vitro anticancer test. Conclusions L-arginase demonstrated more strong anti-cancer cells with an IC50 of 21.4 ug/ml in a dose-dependent manner. L-arginase underwent another investigation for its impact on the caspase 7 and BCL2 family of proteins (BCL2, Bax, and Bax/Bcl2). Through cell arrest in the G1/S phase, L-arginase signals the apoptotic cascade, which is supported by a flow cytometry analysis of cell cycle phases

Rhizopus stolonifer biomass catalytic transesterification capability: optimization of cultivation conditions

Abstract Background Using fungal biomass for biocatalysis is a potential solution for the expensive cost of the use o enzymes. Production of fungal biomass with effective activity requires optimizing the cultivation conditions. Results Rhizopus stolonifer biomass was optimized for transesterification and hydrolysis of waste frying oil (WFO). Growth and biomass lipolytic activities of R. stolonifer improved under shaking conditions compared to static conditions, and 200 rpm was optimum. As biomass lipase and transesterification activities inducer, olive oil was superior to soybean, rapeseed, and waste frying oils. Biomass produced in culture media containing fishmeal as an N-source feedstock had higher lipolytic capabilities than corn-steep liquor and urea. Plackett Burman screening of 9 factors showed that pH (5–9), fishmeal (0.25–1.7%, w/v), and KH2PO4 (0.1–0.9%, w/v) were significant factors with the highest main effect estimates 11.46, 10.42, 14.90, respectively. These factors were selected for response surface methodology (RSM) optimization using central composite design (CCD). CCD models for growth, biomass lipase activity, and transesterification capability were significant. The optimum conditions for growth and lipid modification catalytic activities were pH 7.4, fishmeal (2.62%, w/v), and KH2PO4 (2.99%, w/v). Conclusion Optimized culture conditions improved the whole cell transesterification capability of Rhizopus stolonifer biomass in terms of fatty acid methyl ester (FAME) concentration by 67.65% to a final FAME concentration of 85.5%, w/w

Pathogens Removal in a Sustainable and Economic High-Rate Algal Pond Wastewater Treatment System

This study evaluates the efficiency of a sustainable technology represented in an integrated pilot-scale system, which includes a facultative pond (FP), a high-rate algal pond (HRAP), and a rock filter (RF) for wastewater treatment to produce water that complies with the Egyptian standards for treated wastewater reuse. Still, limited data are available on pathogen removal through HRAP systems. Thus, in this study, the performance of the integrated system was investigated for the removal of Escherichia coli (E. coli), coliform bacteria, eukaryotic pathogens (Cryptosporidium spp., Giardia intestinalis, and helminth ova), somatic coliphages (SOMCPH), and human adenovirus (HAdV). Furthermore, physicochemical parameters were determined in order to evaluate the performance of the integrated system. The principal component analysis and non-metric multidimensional scaling analysis showed a strong significant effect of the integrated system on changing the physicochemical and microbial parameters from inlet to outlet. The mean log10 removal values for total coliform, fecal coliform, and E. coli were 5.67, 5.62, and 5.69, respectively, while 0.88 log10 and 1.65 log10 reductions were observed for HAdV and SOMCPH, respectively. The mean removal of Cryptosporidium spp. and Giardia intestinalis was 0.52 and 2.42 log10, respectively. The integrated system achieved 100% removal of helminth ova. The results demonstrated that the system was able to improve the chemical and microbial characteristics of the outlet to acceptable levels for non-food crops irrigation. Such findings together with low operation and construction costs of HRAPs should facilitate wider implementation of these nature-based systems in remote and rural communities. Overall, this study provides a novel insight into the performance of such systems to eliminate multiple microbial pathogens from wastewater

Insight into antioxidant and anti-inflammatory effects of marine bacterial natural exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats

Abstract Inflammation is a part of the body’s intricate biological reaction to noxious stimuli and defensive reactions. So, the aim of this investigation was to study the anti-inflammatory activity of exopolysaccharide (EPSSM) using carrageenan-induced paw edema in rats. A halophilic bacterial strain was isolated from marine sediments in the Red Sea in Egypt. The isolate has been visually and physiologically recognized, as well as by analyzing its 16S rRNA gene, which confirms Kocuria sp. clone Asker4. This particular isolate can be referenced using the accession number OL798051.1. EPSSM was subjected to purification and fractionation by a DEAE-cellulose column. Preliminary chemical analysis of EPSSM indicated that the monosaccharides were fructose, glucuronic acid, and xylose, with 2.0, 0.5, and 1.0, respectively. The antioxidant potential of EPSSM was investigated, and it was discovered that the level of activity increased independently of the concentrations, reaching a maximum threshold of 94.13% at 100 µg/mL of EPSSM for 120 min. Also, EPSSM at 50 mg/kg orally produced a significant anti-inflammatory effect on the carrageenan model at 2, 3, and 4 intervals. The EPSSM intervention resulted in reductions in the levels of catalase and superoxide dismutase enzymes, as well as a decrease in glutathione. Furthermore, the levels of nitric oxide, lipid peroxidation, and reactive oxygen species resulting from carrageenan-induced edema showed a significant reduction subsequent to the administration of EPSSM. Moreover, the findings indicated that the protein expression levels of cyclooxygenase-2 and interleukin-6 were reduced following treatment with EPSSM, resulting in a reduction of paw edema

Metabolites profiling of <i>Chrysanthemum pacificum</i> Nakai parts using UPLC-PDA-MS coupled to chemometrics

<div><p>Methanol-soluble constituents from the flowers, non-flowering aerial parts and roots of <i>Chrysanthemum pacificum</i> Nakai were analysed <i>via</i> high resolution UPLC-PDA-qTOF-MS followed by chemometrics. Forty-seven chromatographic peaks belonging to various metabolite classes were detected. Most metabolite classes showed qualitative and quantitative differences across parts, with luteolin conjugates being mostly enriched in flowers whereas non-flowering aerial parts contained mostly quercetin and methoxylated flavone conjugates. Root sample ranked the lowest for all flavones and dicaffeoylquinic acids. In contrast, 1,5-di-caffeoylquinic acid levels were found at high levels in flowers and aerial parts reaching 3145 and 1390 μg/g, respectively, suggesting that <i>C. pacificum</i> could serve as a natural resource of this well-recognised anti-hepatotoxic phenolic. Principal component analysis was further used for organs classification in an untargeted manner. This study provides the first map of secondary metabolites distribution in <i>C. pacificum</i> Nakai organs.</p></div