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Molecules (Basel, Switzerland) Mar 2023Crude olive pomace oil (OPO) is a by-product of olive oil extraction. In this study, low-calorie structured triacylglycerols (TAGs) were produced by acidolysis of crude...
Crude olive pomace oil (OPO) is a by-product of olive oil extraction. In this study, low-calorie structured triacylglycerols (TAGs) were produced by acidolysis of crude OPO with medium-chain fatty acids (caprylic, C8:0; capric, C10:0) or interesterification with their ethyl ester forms (C8EE, C10EE). These new TAGs present long-chain fatty acids (L) at position -2 and medium-chain fatty acids (M) at positions -1,3 (MLM). Crude OPO exhibited a high acidity (12.05-28.75% free fatty acids), and high contents of chlorophylls and oxidation products. Reactions were carried out continuously in a packed-bed bioreactor for 70 h, using -1,3 regioselective commercial immobilized lipases ( lipase, Lipozyme TL IM; and lipase, Lipozyme RM IM), in solvent-free media at 40 °C. Lipozyme RM IM presented a higher affinity for C10:0 and C10EE. Lipozyme TL IM preferred C10:0 over C8:0 but C8EE over C10EE. Both biocatalysts showed a high activity and operational stability and were not affected by OPO acidity. The New TAG yields ranged 30-60 and the specific productivity ranged 0.96-1.87 g NewTAG/h.g biocatalyst. Lipozyme RM IM cost is more than seven-fold the Lipozyme TL IM cost. Therefore, using Lipozyme TL IM and crude acidic OPO in a continuous bioreactor will contribute to process sustainability for structured lipid production by lowering the cost of the biocatalyst and avoiding oil refining.
Topics: Olea; Plant Oils; Dietetics; Dietary Fats; Triglycerides; Fatty Acids; Olive Oil; Lipase; Esterification; Enzymes, Immobilized
PubMed: 36985609
DOI: 10.3390/molecules28062637 -
Infection and Drug Resistance 2022Metagenomic next-generation sequencing (mNGS) is a fast, sensitive and accurate diagnostic method for pathogens detection. However, reports on the application of mNGS in...
BACKGROUND
Metagenomic next-generation sequencing (mNGS) is a fast, sensitive and accurate diagnostic method for pathogens detection. However, reports on the application of mNGS in mucormycosis remain scarce.
METHODS
From January 2019 to December 2021, we recruited 13 patients with hematological malignancies who were suspected of mucormycosis and completed mNGS in D20. Then we retrospectively analyze the clinical data, diagnosis, therapeutic process, and outcomes. In order to evaluate the diagnostic value of mNGS in hematological malignancies patients with suspected mucormycosis.
RESULTS
All patients had high risk factors of Invasive Fungal Disease, including hematopoietic stem cell transplantation, immunosuppression, glucocorticoids, etc. The clinical presentations were pulmonary (n=9), rhino-orbito-cerebral (n=4). But the manifestations were nonspecific. All enrolled patients completed mNGS. And most (8/13, 61.54%) of the samples were from blood. Fungi can be detected in all specimens, including Rhizopus (n=7), Rhizomucor (n=4) and Mucor (n=2). In addition, 7/13 (53.85%) specimens were detected bacteria at the same time and virus were detected in 5/13 (38.46%). Histopathological examination was completed in 5 patients, 3 of which were completely consistent with the results of mNGS. After treatment, 6 patients were cured, while the other 7 patients died.
CONCLUSION
mNGS may be a complementary method for early diagnosis, especially for patients who are not suitable for histopathology examination or unable to obtain culture specimen. mNGS can also help detect bacteria and viruses simultaneously, allowing for appropriate and timely antibiotic administration and thus improving patient outcomes.
PubMed: 36570711
DOI: 10.2147/IDR.S393201 -
Current Medical Mycology Mar 2016Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic...
BACKGROUND AND PURPOSE
Soil is the main habitat of saprophytic and pathogenic fungi. Mucoromycotina constitutes a large group of soil fungi, with certain opportunistic members causing systemic infections in immunocompromised hosts. The majority of human and animal infections are caused by the members of the genera , , , (, and . Accordingly, in the present study, we aimed to isolate and identify the main genera of the order Mucorales, using molecular assays and morphological features
MATERIALS AND METHODS
In total, 340 soil samples were collected from seven public parks throughout the city and sidewalk gardens in 14 municipal districts in Isfahan, Iran. All the samples were cultured on the appropriate media, incubated at 27°C for 2- 4 days, and examined daily for visible fungal growth. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was applied and macroscopic, microscopic, and physiological characteristics were assessed to identify fungal colonies.
RESULTS
400 pure colonies, belonging to the orders Mucorales and Mortierellales, including the genera , and were identified The genus (35.5%) was the most frequent isolate, followed by (32.25%) and (27.5%).
CONCLUSION
The results emphasize the importance of opportunistic fungi in public areas and indicate the risk of exposure for immunocompromised individuals.
PubMed: 28681007
DOI: 10.18869/acadpub.cmm.2.1.13 -
Biologia Futura Sep 2021Biogas is the product of anaerobic digestion (AD) of organic waste and is considered to be one of the most valuable natural renewable energy carriers. Plant biomass...
Biogas is the product of anaerobic digestion (AD) of organic waste and is considered to be one of the most valuable natural renewable energy carriers. Plant biomass represents the most abundant eco-friendly energy reservoir on Earth. However, the tenacious and heterogeneous structure of the lignocellulose-rich elements makes it difficult for the involved microbes to digest the recalcitrant substrates. Both the degradation process and the biogas production yield can be enhanced by appropriate pre-treatment of lignocellulosic materials. Filamentous fungi have been known as proficient colonizers of lignocellulosic plant tissues and have been recognized as producers of exceptionally rich and diverse hydrolytic enzymes. We tested Aspergillus nidulans, Trichoderma reesei, Rhizomucor miehei and Gilbertella persicaria filamentous fungal strains for pre-treatment of various agricultural lignocellulosic wastes. During the pre-treatment phase, the β-glucosidase and endoglucanase activity was measured spectrophotometrically. In the AD step, methane production was monitored by gas chromatography. The preliminary results showed that all the applied strains (Aspergillus nidulans, Trichoderma reesei, Rhizomucor miehei and Gilbertella persicaria) were highly effective in producing both β-glucosidase and endo-(1,4)-β-D-glucanase enzymes, which might explain the greatly improved AD results. Pre-treatment with the above-mentioned filamentous fungi positively affected the biogas production, although the effect strongly depended on the selection of the fungal partner for any given biomass substrate. Depending on the used substrate and the pre-treatment strain, overall methane yields were elevated two-fold relative to the controls.
Topics: Agrochemicals; Biofuels; Fungi; Industrial Waste
PubMed: 34554554
DOI: 10.1007/s42977-021-00083-3 -
FEMS Microbiology Ecology Sep 2020Rhizopus arrhizus is a common pathogenic Mucoralean mold that exists as a saprophyte, and is disseminated through sporangiospores, which germinate to form mycelia under...
Rhizopus arrhizus is a common pathogenic Mucoralean mold that exists as a saprophyte, and is disseminated through sporangiospores, which germinate to form mycelia under suitable environmental or infection settings. Such morphological transitions are often mediated by self-produced effector molecules in a density-dependent fashion. This study aimed to elucidate if a quorum-dependent, cell-density-driven phenomenon exists in R. arrhizus, and identify the molecule(s) involved. The germination of R. arrhizus was observed to be reliant on the seeding density, with nearly 71% and 47% germination in Sabouraud dextrose and glucose asparagine media respectively at 1 × 105-1 × 106 spores/mL, and only 10% and 1% germination respectively with 1 × 108 spores/mL. The late-growth-stage supernatant also hindered the spore germination and liquid-culture biomass in a dose-dependent way. These effects were being mediated by a volatile inhibitor present in the headspace and supernatant of R. arrhizus cultures, identified as 2-methyl-2-butene by gas chromatography and electron ionization-quadrupole mass spectrometry. The compound was present in a density-dependent manner and considerably impaired fungal germ-tube emergence and elongation during germination. Spore swelling remained unaffected. Multiple thin protrusions comprising of F-actin and microtubules were seen emanating from the treated cells, suggestive of filopodia-like and cytoneme-like extensions. The same compound was also detected in Rhizomucor pusillus.
Topics: Culture Media; Rhizomucor; Rhizopus; Rhizopus oryzae; Spores, Fungal
PubMed: 32816006
DOI: 10.1093/femsec/fiaa170 -
European Journal of Pharmaceutics and... Feb 2022High throughput screening for measuring the stability of industrially relevant proteins and their variants is necessary for quality assessment in the development...
High throughput screening for measuring the stability of industrially relevant proteins and their variants is necessary for quality assessment in the development process. Advances in automation, measurement time and sample consumption for many techniques allow rapid measurements with minimal amount of protein. However, many methods include automated data analysis, potentially neglecting important aspects of the protein's behavior in certain conditions. In this study we implement small angle X-ray scattering (SAXS), typically not used to assess protein behavior in industrial screening, in a high throughput screening workflow to address problems of contradicting results and reproducibility among different high throughput methods. As a case study we use the lipases of Thermomyces lanuginosus and Rhizomucor miehei, widely used industrial biocatalysts. We show that even the initial analysis of the SAXS data without performing any time-consuming modelling provide valuable information on interparticle interactions. We conclude that recent advances in automation and data processing, have enabled SAXS to be used more widely as a tool to gain in-depth knowledge highly useful for protein formulation development. This is especially relevant in light of increasing accessibility to SAXS due to the commercial availability of benchtop instruments.
Topics: High-Throughput Screening Assays; Humans; Protein Stability; Proteins; Reproducibility of Results; Scattering, Small Angle; X-Ray Diffraction
PubMed: 34826593
DOI: 10.1016/j.ejpb.2021.08.018 -
Journal of Chemical Information and... Nov 2019Most natural proteins exhibit poor thermostability, which limits their industrial application. Computer-aided rational design is an efficient purpose-oriented method...
Most natural proteins exhibit poor thermostability, which limits their industrial application. Computer-aided rational design is an efficient purpose-oriented method that can improve protein thermostability. Numerous machine-learning-based methods have been designed to predict the changes in protein thermostability induced by mutations. However, all of these methods have certain limitations due to existing mutation coding methods that overlook protein sequence features. Here we propose a method to predict protein thermostability using convolutional neural networks based on an in-depth study of thermostability-related protein properties. This method comprises a three-dimensional coding algorithm, including protein mutation information and a strategy to extract neighboring features at protein mutation sites based on multiscale convolution. The accuracies on the S1615 and S388 data sets, which are widely used for protein thermostability predictions, reached 86.4 and 87%, respectively. The Matthews correlation coefficient was nearly double those produced using other methods. Furthermore, a model was constructed to predict the thermostability of lipase mutants based on the S3661 data set, a single amino acid mutation data set screened from the ProTherm protein thermodynamics database. Compared with the RIF strategy, which consists of three algorithms, i.e., Rosetta ddg monomer, I Mutant 3.0, and FoldX, the accuracy of the proposed method was higher (75.0 vs 66.7%), and the negative sample resolution was simultaneously enhanced. These results indicate that our prediction method more effectively assessed the protein thermostability and distinguished its features, making it a powerful tool to devise mutations that enhance the thermostability of proteins, particularly enzymes.
Topics: Animals; Humans; Models, Chemical; Models, Molecular; Neural Networks, Computer; Point Mutation; Protein Stability; Proteins; Temperature; Thermodynamics
PubMed: 31657922
DOI: 10.1021/acs.jcim.9b00220 -
Transplant Infectious Disease : An... Jun 2021Individuals diagnosed with acute lymphoid and myeloid malignancies are at significant risk of invasive fungal and bacterial infections secondary to their marked... (Review)
Review
The role of 18F-Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography (FDG PET/CT) in assessment of complex invasive fungal disease and opportunistic co-infections in patients with acute leukemia prior to allogeneic hematopoietic cell transplant.
INTRODUCTION
Individuals diagnosed with acute lymphoid and myeloid malignancies are at significant risk of invasive fungal and bacterial infections secondary to their marked immunocompromised states with a significant high risk of mortality. The role of metabolic imaging with 18F-Fluorodeoxyglucose (FDG) Positron Emission Tomography/Computed Tomography (PET/CT) has been increasingly recognized in optimizing the diagnosis of invasive infection, monitoring the response to therapy and guiding the duration of antimicrobial therapy or need to escalate to surgical intervention.
METHODS
Two distinct cases of pulmonary co-infection of rare fungal and bacterial pathogens are explored in severely immunocompromised individuals where FDG PET/CT aided both patients to make a full recovery and transition to HCT. The first case explores mixed Scedosporium apiospermum and Rhizomucor pulmonary infection on a background of T cell/myeloid mixed phenotype acute leukemia ultimately warranting long-term antifungal therapy and lobectomy prior to HCT. The second case explores Fusarium and Nocardia pulmonary infection on a background of relapsed AML also warranting surgical resection with lobectomy and long-term antimicrobials prior to transition to HCT.
DISCUSSION
The cases highlight the utility of FDG PET/CT to support the diagnosis of infections, including the presence or absence of disseminated infection, and to provide highly sensitive monitoring of the infection over time. FDG PET/CT played a key role in directing therapy duration decisions and prompted the necessity for surgical intervention. Ultimately, the use of FDG PET/CT allowed for a successful transition to HCT highlighting its value in this clinical setting.
CONCLUSION
FDG PET/CT has an emerging role in the diagnostic and monitoring pathway for complex infections in high-risk immunocompromised patients.
Topics: Coinfection; Fluorodeoxyglucose F18; Hematopoietic Stem Cell Transplantation; Humans; Invasive Fungal Infections; Leukemia, Myeloid, Acute; Positron Emission Tomography Computed Tomography; Positron-Emission Tomography; Radiopharmaceuticals; Transplantation, Homologous
PubMed: 33338319
DOI: 10.1111/tid.13547 -
Revista Iberoamericana de Micologia 2015Mucormycosis is usually an acute angioinvasive infections, which leads to non-suppurative necrosis and significant tissue damage. It represents 1.6% of all the invasive... (Review)
Review
Mucormycosis is usually an acute angioinvasive infections, which leads to non-suppurative necrosis and significant tissue damage. It represents 1.6% of all the invasive fungal infections and predominates in immunosuppressed patients with risk factors. Incidence has been significantly increased even in immunocompetent patients. Due to finding a case of disseminated mucormycosis caused by Rhizomucor pusillus in a young immunocompetent patient, a systematic review was carried out of reported cases in PubMed of mucormycosis in immunocompetent adults according to the main anatomic locations, and especially in disseminated cases. A review of the main risk factors and pathogenicity, clinical manifestations, techniques of early diagnosis, current treatment options, and prognosis is presented. Taxonomy and classification of the genus Mucor has also been reviewed.
Topics: Agricultural Workers' Diseases; Antifungal Agents; Combined Modality Therapy; Dermatomycoses; Fungemia; Fungi; Humans; Immunocompetence; Incidence; Mucormycosis; Opportunistic Infections; Respiratory Tract Infections; Risk Factors; Soil Microbiology
PubMed: 25543322
DOI: 10.1016/j.riam.2014.01.006 -
Soft Matter Aug 2021The use of enzymes as biocatalysts in industrial applications has received much attention during the last few years. Lipases are widely employed in the food and cosmetic...
The use of enzymes as biocatalysts in industrial applications has received much attention during the last few years. Lipases are widely employed in the food and cosmetic industry, for the synthesis of novel biomaterials and as a greener solution for the treatment of waste cooking oils (WCO). The latter topic has been widely explored with the use of enzymes from several origins and types, for the treatment of different used and non-used cooking oils. The experimental conditions of such works are also quite broad, hampering the detailed understanding of the process. In this work we present a detailed characterization of the interaction of several commonly used lipases with different types of vegetal oils and food fats through coarse-grained molecular dynamics simulations. First, the molecular details of the oil/water (O/W) mixtures, namely at the O/W interface, are described. The O/W interface was found to be enriched in triglyceride molecules with higher polarity. Then, the interaction of lipases with oil mixtures is characterized from different perspectives, including the identification of the most important protein residues for this process. The lipases from Thermomyces lanuginosus (TLL), Rhizomucor miehei (RML) and Candida antarctica (CALB) were found to bind to the O/W interface in a manner that makes the protein binding site more available for the oil molecules. These enzymes were also found to efficiently bind to the O/W interface of all oil mixtures, which in addition to reactivity factors, may explain the efficient applicability of these enzymes to a large variety of edible oils and WCO.
Topics: Basidiomycota; Enzymes, Immobilized; Eurotiales; Oils; Rhizomucor; Water
PubMed: 34155497
DOI: 10.1039/d1sm00590a