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Beta-mannans are insoluble plant polysaccharides with beta-1,4-linked mannose as the backbone. We used three forms of this polysaccharide, namely, pure mannan, glucomannan, and galactomannan, to enrich haloarchaea, which have the ability to utilize mannans for growth. Four mannan-utilizing strains obtained in pure cultures were closely related to each other on the level of the same species. Furthermore, another strain selected from the same habitats with a soluble beta-1,4-glucan (xyloglucan) was also able to grow with mannan. The phylogenomic analysis placed the isolates into a separate lineage of the new genus level within the family Natrialbaceae of the class Halobacteria. The strains are moderate alkaliphiles, extremely halophilic, and aerobic saccharolytics. In addition to the three beta-mannan forms, they can also grow with cellulose, xylan, and xyloglucan. Functional genome analysis of two representative strains demonstrated the presence of several genes coding for extracellular endo-beta-1,4-mannanase from the GH5_7 and 5_8 subfamilies and the GH26 family of glycosyl hydrolases. Furthermore, a large spectrum of genes encoding other glycoside hydrolases that were potentially involved in the hydrolysis of cellulose and xylan were also identified in the genomes. A comparative genomics analysis also showed the presence of similar endo-beta-1,4-mannanase homologs in the cellulotrophic genera Natronobiforma and Halococcoides. Based on the unique physiological properties and the results of phylogenomic analysis, the novel mannan-utilizing halolarchaea are proposed to be classified into a new genus and species Natronoglomus mannanivorans gen. nov., sp. nov. with the type strain AArc-m2/3/4 (=JCM 34861=UQM 41565)., BT/Environmental Biotechnology

The optimal cut-off value of the optical density index of the galactomannan antigen assays (GM) for diagnosing invasive pulmonary aspergillosis in hematological patients is a disputed topic. This article conducts a systematic review with a meta-analysis to establish which optical density index (ODI) cut-off value should be implemented into clinical practice. Pubmed, Embase and Cochrane databases were searched (N = 27). The pooled data, using a generalized linear mixed model with binomial distribution, resulted in an overall serum sensitivity of 0.76 and a specificity of 0.92. For serum ODI 0.5 there was a pooled sensitivity of 0.92 and a specificity of 0.84. The pooled data of all broncho-alveolar lavage (BAL) studies resulted in an overall sensitivity of 0.80 and a specificity of 0.95. For BAL ODI 0.5, there was a pooled sensitivity of 0.75 and a specificity of 0.88. For the BAL ODI 1.0 pooling, the studies resulted in a sensitivity of 0.75 and a specificity of 0.96. Serum ODI of 0.5 and BAL ODI of 1.0 are the most suitable cut-offs for clinical practice. However, our study affirms that the evidence for the use of GM in clinical practice for the hematological malignancy patient is currently insufficient and more research is needed to determine the diagnostic value of GM.

This multicenter study evaluated the IMMY Aspergillus Galactomannan Lateral Flow Assay (LFA) with automated reader for diagnosis of pulmonary aspergillosis in patients with COVID-19-associated acute respiratory failure (ARF) requiring intensive care unit (ICU) admission between 03/2020 and 04/2021. A total of 196 respiratory samples and 148 serum samples (n = 344) from 238 patients were retrospectively included, with a maximum of one of each sample type per patient. Cases were retrospectively classified for COVID-19-associated pulmonary aspergillosis (CAPA) status following the 2020 consensus criteria, with the exclusion of LFA results as a mycological criterion. At the 1.0 cutoff, sensitivity of LFA for CAPA (proven/probable/possible) was 52%, 80% and 81%, and specificity was 98%, 88% and 67%, for bronchoalveolar lavage fluid (BALF), nondirected bronchoalveolar lavage (NBL), and tracheal aspiration (TA), respectively. At the 0.5 manufacturer's cutoff, sensitivity was 72%, 90% and 100%, and specificity was 79%, 83% and 44%, for BALF, NBL and TA, respectively. When combining all respiratory samples, the receiver operating characteristic (ROC) area under the curve (AUC) was 0.823, versus 0.754, 0.890 and 0.814 for BALF, NBL and TA, respectively. Sensitivity and specificity of serum LFA were 20% and 93%, respectively, at the 0.5 ODI cutoff. Overall, the Aspergillus Galactomannan LFA showed good performances for CAPA diagnosis, when used from respiratory samples at the 1.0 cutoff, while sensitivity from serum was limited, linked to weak invasiveness during CAPA. As some false-positive results can occur, isolated results slightly above the recommended cutoff should lead to further mycological investigations.

Critically ill patients with coronavirus disease 2019 (COVID-19) may develop COVID-19-associated pulmonary aspergillosis (CAPA), which impacts their chances of survival. Whether positive bronchoalveolar lavage fluid (BALF) mycological tests can be used as a survival proxy remains unknown. We conducted a post hoc analysis of a previous multicenter, multinational observational study with the aim of assessing the differential prognostic impact of BALF mycological tests, namely, positive (optical density index of ≥1.0) BALF galactomannan (GM) and positive BALF Aspergillus culture alone or in combination for critically ill patients with COVID-19. Of the 592 critically ill patients with COVID-19 enrolled in the main study, 218 were included in this post hoc analysis, as they had both test results available. CAPA was diagnosed in 56/218 patients (26%). Most cases were probable CAPA (51/56 [91%]) and fewer were proven CAPA (5/56 [9%]). In the final multivariable model adjusted for between-center heterogeneity, an independent association with 90-day mortality was observed for the combination of positive BALF GM and positive BALF Aspergillus culture in comparison with both tests negative (hazard ratio, 2.53; 95% CI confidence interval [CI], 1.28 to 5.02; P = 0.008). The other independent predictors of 90-day mortality were increasing age and active malignant disease. In conclusion, the combination of positive BALF GM and positive BALF Aspergillus culture was associated with increased 90-day mortality in critically ill patients with COVID-19. Additional study is needed to explore the possible prognostic value of other BALF markers.

Invasive pulmonary aspergillosis (IPA) is a life-threatening disease that affects mainly immunocompromised hosts. Galactomannan testing from serum and bronchoalveolar lavage fluid (BALF) represents a cornerstone in diagnosing the disease. Here, we evaluated the diagnostic performance of the novel Aspergillus-specific galactomannoprotein (GP) enzyme-linked immunosorbent assay (ELISA; Euroimmun Medizinische Labordiagnostika) compared with the established Platelia Aspergillus GM ELISA (GM; Bio-Rad Laboratories) for the detection of Aspergillus antigen in BALF. Using the GP ELISA, we retrospectively tested 115 BALF samples from 115 patients with clinical suspicion of IPA and GM analysis ordered in clinical routine. Spearman's correlation statistics and receiver operating characteristics (ROC) curve analysis were performed. Optimal cutoff values were determined using Youden's index. Of 115 patients, 1 patient fulfilled criteria for proven IPA, 42 for probable IPA, 15 for putative IPA, 10 for possible IPA, and 47 did not meet criteria for IPA. Sensitivities and specificities for differentiating proven/probable/putative versus no IPA (possible excluded) were 74% and 96% for BALF GP and 90% and 96% for BALF GM at the manufacturer-recommended cutoffs. Using the calculated optimal cutoff value of 12 pg/mL, sensitivity and specificity of the BALF GP were 90% and 96%, respectively. ROC curve analysis showed an area under the curve (AUC) of 0.959 (95% confidence interval [CI] of 0.923 to 0.995) for the GP ELISA and an AUC of 0.960 (95% CI of 0.921 to 0.999) for the GM ELISA for differentiating proven/probable/putative IPA versus no IPA. Spearman's correlation analysis showed a strong correlation between the ELISAs (rho = 0.809, P < 0.0001). The GP ELISA demonstrated strong correlation and test performance similar to that of the GM ELISA and could serve as an alternative test for BALF from patients at risk for IPA.

Carbohydrate active enzymes, such as those involved in plant cell wall and storage polysaccharide biosynthesis and deconstruction, often contain repeating non-catalytic carbohydrate binding modules (CBMs) to compensate for low-affinity binding typical of protein-carbohydrate interactions. The bacterium Saccharophagus degradans produces an endo-β-mannanase of glycoside hydrolase family 5 subfamily 8 with three phylogenetically distinct family 10 CBMs located C-terminally from the catalytic domain (SdGH5_8-CBM10x3). However, the functional roles and cooperativity of these CBM domains in polysaccharide binding is not clear. To learn more we studied the full-length enzyme, three stepwise CBM10 truncations, and green fluorescent protein fusions of the individual CBM10s and all three domains together by pull-down assays, affinity gel electrophoresis, and activity assays. Only the C-terminal CBM10-3 was found to bind strongly to microcrystalline cellulose (dissociation constant, Kd = 1.48 μM). CBM10-3 and CBM10-2 bound galactomannan with similar affinity (Kd = 0.2-0.4 mg/ml), but CBM10-1 had 20-fold lower affinity for this substrate. CBM10 truncations barely affected specific activity on carob galactomannan and konjac glucomannan. Full-length SdGH5_8-CBM10x3 was two-fold more active on the highly galactose-decorated viscous guar gum galactomannan and crystalline ivory nut mannan at high enzyme concentrations, but the specific activity was 4- to 9-fold reduced at low enzyme and substrate concentrations compared to the enzyme lacking CBM10-2 and -3. Comparison of activity and binding data for the different enzyme forms indicates unproductive and productive polysaccharide binding to occur. We conclude that the C-terminal-most CBM10-3 secures firm binding, with contribution from CBM10-2, which with CBM10-1 also provides spatial flexibility.

Invasive aspergillosis (IA) is an increasingly recognised phenomenon in critically ill patients in the intensive care unit, including in patients with severe influenza and severe coronavirus disease 2019 (COVID-19) infection. To date, there are no consensus criteria on how to define IA in the ICU population, although several criteria are used, including the AspICU criteria and new consensus criteria to categorise COVID-19-associated pulmonary aspergillosis (CAPA). In this review, we describe the epidemiology of IA in critically ill patients, most common definitions used to define IA in this population, and most common clinical specimens obtained for establishing a mycological diagnosis of IA in the critically ill. We also review the most common diagnostic tests used to diagnose IA in this population, and lastly discuss the most common clinical presentation and imaging findings of IA in the critically ill and discuss areas of further needed investigation.

[EN] A fenugreek seed gum, extracted fromTrigonella foenum-graecumseeds and rich in galactomannan, was chemically and physically characterised and its prebiotic potential for young rabbits was evaluated in vitro, both as pure fenugreek seed gum and when included up to 20 g/kg in rabbit diets rich in soluble and insoluble fibre. Fenugreek seed gum was resistant to pepsin and pancreatin digestion but was totally fermented by rabbit caecal bacteria. Fenugreek seed gum linear inclusion up to 20 g/kg in diets rich in soluble fibre has led to a reduction in the solubility of some nutrients during in vitro enzymatic phase and an increase in the fermented fraction. Fenugreek seed gum satisfies two essential conditions of a prebiotic: resistance to enzymatic digestion and being totally fermented by caecal bacteria. Some components of soluble fibre appear to have prebiotic effects that can contribute to improving digestive health in post-weaning rabbits. In this work, a fenugreek seed gum (FGS), extracted fromTrigonella foenum-graecumseeds and rich in galactomannan, was characterised. Both the pure FSG and ten substrates obtained by the inclusion of 0, 5, 10, 15 and 20 g/kg of FSG in diets rich in soluble (SF) and insoluble (IF) fibre were evaluated in vitro to determine FSG prebiotic potential for rabbit diets. FSG was rich in total sugars (630 g/kg dry matter), consisting entirely of galactose and mannose in a 1:1 ratio, and a moderate protein content (223 g/kg dry matter). Pure FSG was affected very little by in vitro digestion, as only 145 g/kg of the FSG was dissolved during the enzymatic phase. However, the linear inclusion of FSG up to 20 g/kg in growing rabbit feeds has led to a reduction in the solubility of some nutrients during in vitro enzymatic phase, especially in SF diets. Pure FSG not digested during the enzymatic phase almost completely disappeared during the in vitro fermentation phase, 984 g/kg of this indigestible fraction. However, although linear inclusi

Background: Invasive pulmonary aspergillosis (IPA) is an increasingly recognized complication in intensive care unit (ICU) patients, especially those with infuenza, cirrhosis, chronic obstructive pulmonary disease, and other dise

[ES] Esta tesis aborda la caracterización y posible utilización de la goma de alholva (Trigonella foenum-graecum) (FSG, por sus siglas en inglés) en la nutrición de gazapos durante el período posterior al destete como una alternativa al uso de antimicrobianos, centrándose en el comportamiento de la goma extraída con diferentes niveles de fibra dietaria, tanto in vitro como in vivo. En el primer experimento, a partir de una dieta basal comercial de conejo, se formularon tres dietas experimentales con niveles graduales de inclusión de FSG (0, 0.25, 0.50%) administrados a tres grupos de conejos desde el destete a los 31 días hasta el sacrificio a los 94 días. Se estudió el efecto de las dietas experimentales sobre la digestibilidad fecal en dos edades (38-41 días y 56-59 días), además de los parámetros de fermentación cecal al final del experimento. Por otro lado, a partir de la misma dieta comercial de conejo, se formularon cinco dietas experimentales que contenían 0. 0.125, 0.25, 0.50 y 100% de FSG, que se sometieron a incubación in vitro con inóculo cecal de conejo durante 72 h. Se midieron la producción de gas y las variables de fermentación. La digestibilidad fecal aparente y los parámetros de fermentación cecal no se vieron afectados significativamente por la inclusión in vivo de FSG hasta 0.5%. Sin embargo, los animales alimentados con FSG mostraron valores de pH cecal más bajos. La inclusión gradual en la dieta de FSG aumentó la concentración in vitro de ácidos grasos volátiles (VFA, por sus siglas en inglés), mientras que el FSG puro aumentó la producción asintótica de gas y la tasa máxima de degradación del sustrato y disminuyó el tiempo de incubación en el que se forma la mitad de la cantidad asintótica de gas. La incubación in vitro de FSG puro disminuyó el valor del pH, la concentración de ácido láctico y la concentración de N-NH3, y aumentó la de VFA. En el segundo experimento, el FSG se caracterizó para determinar su composición química, su contenido de, [EN] This thesis tackles the possible characterisation and utilisation of fenugreek (Trigonella foenum-graecum) seed gum (FSG) in the nutrition of young rabbits during the post-weaning period as an alternative to antimicrobials uses. The study was focussed on the behaviour of the extracted gum with different dietary fibre levels, both in vitro and in vivo. In a first trial, starting from a basal commercial rabbit diet, three experimental diets were formulated with gradual levels of inclusion of FSG (0, 0.25, 0.50 %) given to three groups of rabbits starting from weaning at 31 days old to slaughter 94 days old. The effect of the experimental diets was studied on the faecal digestibility of the diet in two ages (38-41 days and 56-59days) and on the caecal fermentation parameters at the end of the experiment. On the other hand, starting from the same commercial rabbit diet five experimental diets were formulated containing 0, 0.125, 0.25, 0.50 and 100% of FSG that were submitted to in vitro incubation with rabbit caecal inoculum during 72h. Gas production and fermentation traits were measured. Apparent faecal digestibility and caecal fermentation parameters were not significantly affected by the in vivo inclusion of FSG up to 0.5%. However, animals fed with FSG showed lower caecal pH values. Gradual dietary inclusion of FSG increased in vitro concentration of volatile fatty acids (VFA), while pure FSG increased the asymptotic gas production and the maximum substrate degradation rate and decreased the time after incubation at which half of the asymptotic amount of gas is formed. In vitro incubation of pure FSG decreased pH value, lactic acid concentration and N-NH3 concentration and increased that of VFA. In a second trial, FSG was characterised determining its chemical composition, galactose and mannose content and prebiotic potential. Pure FSG and gradual levels of FSG (0, 5, 10, 15 and 20 g/kg), included both in diets rich in soluble fibre (SF) from beet pulp (includ, [CA] Aquesta tesi aborda la caracterització i possible utilització de la goma de fenigrec (Trigonella foenum-graecum) (FSG, per les sigles en anglès) en la nutrició del conills durant el període posterior al deslletament com una alternativa a l'ús d'antimicrobians, centrant-se en el comportament de la goma extreta amb diferents nivells de fibra dietària, tant in vitro com in vivo. En el primer experiment, a partir d'una dieta basal comercial de conill, es van formular tres dietes experimentals amb nivells graduals d'inclusió de FSG (0, 0.25, 0.50%) administrats a tres grups de conills des del deslletament als 31 dies fins al sacrifici als 94 dies. Es va estudiar l'efecte de les dietes experimentals sobre la digestibilitat fecal en dues edats (38-41 dies i 56-59 dies), a més dels paràmetres de fermentació cecal a la fi de l'experiment. D'altra banda, a partir de la mateixa dieta comercial de conill, es van formular cinq dietes experimentals que contenien 0. 0.125, 0.25, 0,50 i 100% de FSG, que es van sotmetre a incubació in vitro amb inòcul cecal de conill durant 72 h. Es van mesurar la producció de gas i les variables de fermentació. La digestibilitat fecal aparent i els paràmetres de fermentació cecal no es van veure afectats significativament per la inclusió in vivo de FSG fins a 0.5%. No obstant això, els animals alimentats amb FSG van mostrar valors de pH cecal més baixos. La inclusió gradual en la dieta de FSG va augmentar la concentració in vitro d'àcids grassos volàtils (VFA, per les sigles en anglès), mentre que el FSG pur va augmentar la producció asimptòtica de gas i la taxa màxima de degradació del substrat i va disminuir el temps d'incubació en el qual es forma la meitat de la quantitat asimptòtica de gas. La incubació in vitro d'FSG pur va disminuir el valor del pH, la concentració d'àcid làctic i la concentració de N-NH3, i va augmentar la de VFA. En el segon experiment, el FSG es va caracteritzar per determinar la seva composició química, el seu conti

Galactomannan (GM) detection in biological samples has been shown to predict therapeutic response by azoles and polyenes. In a murine invasive pulmonary aspergillosis model, fosmanogepix or posaconazole treatment resulted in an ∼6- to 7-log reduction in conidial equivalents (CE)/g lung tissue after 96 h versus placebo. Changes in GM levels in BAL fluid and serum mirrored reductions in lung CE, with significant decreases seen after 96 h or 72 h for fosmanogepix or posaconazole, respectively (P < 0.02).

[ES] En el presente trabajo se ha estudiado el efecto de la incorporación de harina de fenogreco (2,5, 5, 7,5 y 10% g/g sólidos) para la formulación de pasta fresca a base de harina de garbanzo y de chufa. En concreto, se han evaluado las propiedades reológicas (test oscilatorio y capacitancia de la fluencia), la humedad y la microestructura de las masas obtenidas. Las harinas empleadas fueron caracterizadas en términos de distribución de tamaño de partícula, propiedades funcionales (capacidad de retención de agua y capacidad de absorción de grasa) y microestructura. La incorporación de fenogreco supuso un aumento de la capacidad de retención de agua debido a la presencia de galactomanano, pero no influyó en la capacidad de absorción de grasa. Los resultados referentes a la reología de la masa revelaron un progresivo aumento del módulo de almacenamiento y del módulo de pérdidas, así como una reducción de la capacitancia de la fluencia, con el incremento de fenogreco en las formulaciones. Por tanto, la estructura de las masas se vuelve más resistente a la deformación y con un carácter más semisólido. El análisis de microscopía permitió observar el efecto del galactomanano en el desarrollo de una matriz más densa con los gránulos de almidón embebidos en ella y rodeados por el mucílago de la fibra soluble hidratada. Tras la cocción, las micrografías revelaron una menor gelatinización del almidón a altos porcentajes de fenogreco, probablemente debido la mayor competencia por el agua entre la fibra soluble y el almidón., [EN] In this study, it was assessed the effect of incorporating fenugreek (2,5, 5, 7,5 and 10% g/g solids) for the formulation of fresh pasta based on chickpea flour and tiger nut flour. Concretely, it have been evaluated the rheological properties (oscillatory test and creep test), water content and microstructure of the obtained doughs. Flours were characterized in terms of particle size distribution, functional properties (water holding capacity and fat absorption capacity) and microstructure. The incorporation of fenugreek resulted in an increase of the water holding capacity due to the presence of galactomannan, but fat absorption capacity was not influenced. Concerning dough rheology, results revealed a progressive increase of the storage modulus and the loss modulus, as well as a reduction of the instant capacitance, due to the increase of fenugreek in the formulations. Therefore, dough structure becomes more resistant to deformation and with a more semi-solid character. The microscopy analysis allowed observing the effect of the galactomannan presence in the development of a denser matrix with the starch granules embedded in it and surrounded by the mucilage of the hydrated soluble fiber. After cooking, the micrographs revealed a lower gelatinization of the starch at high percentages of fenugreek, probably due to the higher competition for water between the soluble fiber and the starch.

BackgroundWe compared new Aspergillus Galactomannan Lateral Flow Assay with the newly formatted Aspergillus-specific Lateral Flow device tests for the diagnosis of invasive pulmonary aspergillosis (IPA) in non-neutropenic patients.MethodsWe performed both tests in 82 bronchoalveolar lavage fluid samples from 82 patients at risk for IPA but without underlying haematologic malignancy. Samples were collected between September 2016 and September 2018 at the University of California San Diego, United States. IPA was classified following two published consensus criteria.ResultsClassification of cases varied widely between the two consensus criteria. When using criteria established for the intensive care unit, 26/82 patients (32%) met criteria for proven or putative IPA. Both point-of-care assays showed sensitivities ranging between 58% and 69%, with specificities between 68% and 75%. Sensitivity increased up to 81% when both tests were combined.ConclusionThe study outlines the need for updated, unified and more broadly applicable consensus definitions for classifying IPA in non-neutropenic patients, a work that is currently in progress. Both point-of-care tests showed comparable performance, with sensitivities and specificities in the 60%-70% range when used alone and increasing to 80% when used in combination. The new point-of-care tests may serve a role at the bedside in those with clinical suspicion of IPA.

Aerogels are obtained by laccase/2,2,6,6-tetramethylpiperidin-1-yl)oxyl (TEMPO)-oxidation of galactomannans (GMs) from the leguminous plants fenugreek (Trigonella foenum-graecum), sesbania (Sesbania bispinosa) and guar (Cyamopsis tetragonolobus). GM oxidation in aqueous solutions causes a viscosity increase, resulting in structured and stable hydrogels. Upon lyophilization, water-insoluble aerogels are obtained, capable of water uptake up to several times their own weight. The materials derived from the three gums have been analyzed and compared. Chemical modifications have been studied by electrospray-ionization mass spectrometry (ESI-MS) and Fourier-transform infrared spectroscopy (FTIR). Polymer structure has been determined by liquid-state and semi-quantitative solid-state nuclear magnetic resonance (NMR) spectroscopy and chemical stability by incubation under variable conditions of pH, ionic strength and solvent nature. The results show that hydrogel formation is due to oxidation of primary hydroxyl groups to carbonyl and carboxyl groups and subsequent formation of hemiacetal and ester bonds. Fenugreek displays the highest stability, compared to guar and sesbania rehydrated aerogels. This feature could be interpreted by its higher degree of substitution (Gal:Man = 1:1) and consequently higher amount of galactose primary alcohols, leading to more extensive crosslinking

Background Invasive pulmonary aspergillosis typically occurs in an immunocompromised host. For almost a century, influenza has been known to set up for bacterial superinfections, but recently patients with severe influenza were also reported to develop invasive pulmonary aspergillosis. We aimed to measure the incidence of invasive pulmonary aspergillosis over several seasons in patients with influenza pneumonia in the intensive care unit (ICU) and to assess whether influenza was an independent risk factor for invasive pulmonary aspergillosis. Methods We did a retrospective multicentre cohort study. Data were collected from adult patients with severe influenza admitted to seven ICUs across Belgium and The Netherlands during seven influenza seasons. Patients were older than 18 years, were admitted to the ICU for more than 24 h with acute respiratory failure, had pulmonary infiltrates on imaging, and a confirmed influenza infection based on a positive airway PCR test (influenza cohort). We used logistic regression analyses to determine if influenza was independently associated with invasive pulmonary aspergillosis in non-immunocompromised (ie, no European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group [EORTC/MSG] host factor) influenza-positive patients (influenza case group) compared with nonimmunocompromised patients with severe community-acquired pneumonia who had a negative airway influenza PCR test (control group). Findings Data were collected from patients admitted to the ICU between Jan 1, 2009, and June 30, 2016. Invasive pulmonary aspergillosis was diagnosed in 83 (19%) of 432 patients admitted with influenza (influenza cohort), a median of 3 days after admission to the ICU. The incidence was similar for influenza A and B. For patients with influenza who were immunocompromised, incidence of invasive pulmonary aspergillosis was as hi

Aerogels were obtained from laccase/TEMPO-oxidized galactomannans (GM) of the leguminous plants: fenugreek, sesbania and guar, with several potential applications, such as biocompatible, biodegradable “delivery systems” (DS) of active principles. Chemo-enzymatic oxidation of GM in aqueous solution caused a viscosity increase up to fifteen-fold, generating structured, elastic, and stable hydrogels, presumably due to formation of carbonyl groups from primary OH's on the polymers and subsequent establishment of hemiacetalic bonds with available free hydroxyl groups. Upon lyophilization of these hydrogels, water-insoluble aerogels were generated, capable of uptaking an aqueous solution several times their own weight. In the present paper, these aerogels were further characterized by ESI-MS after enzymatic depolymerization, scanning electron microscopy, thermal and X-ray analyses. Fenugreek confirmed to be more structured and stable compared to guar and sesbania aerogels, which is likely due to its higher amount of oxidizable galactose units linked to the mannose backbone (i.e., Gal: Man = 1:1) and, therefore, to more extensive cross-linking of the resulting elastic gel.

We describe aerogels obtained by laccase/TEMPO-oxidation and lyophilization of galactomannans (GM) from fenugreek, sesbania and guar. Enzymatic oxidation of GM in aqueous solution caused viscosity increase up to fifteen-fold, generating structured, elastic, stable hydrogels, presumably due to establishment of hemiacetalic bonds between newly formed carbonyl groups and free hydroxyl groups. Upon lyophilization, water-insoluble aerogels were obtained, whose mechanical properties and porosity were investigated. Active principles were absorbed into the aerogels from aqueous solutions and, following rinsing, blotting, re-lyophilization, were released in an appropriate medium. The release of the antibiotic polymyxin B was tested against six different Gram-negative bacterial strains, of the antimicrobial peptide nisin against two Gram-positive and of the muraminidase lysozyme against one anaerobic strain. Protease and lipase release in solution from “enzyme loaded” aerogels was monitored by the increase in enzymatic activity. These biomaterials could represent new versatile, biocompatible “delivery systems” of actives for biomedical and industrial applications

This paper reports the preparation of galactomannan/ionic liquid composite materials from the corresponding ion gels. Three kinds of galactomannans, that is, fenugreek gum (FG), guar gum (GG), and locust bean gum (LBG) and an ionic liquid of 1-butyl-3-methylimidazolium chloride (BMIMCl) were used. When the galactomannan/BMIMCl gels were immersed in ethanol, followed by dryness under reduced pressure, the galactomannan/BMIMCl composite materials were obtained. The crystalline structures of galactomannans in the materials were evaluated by the powder X-ray diffraction measurement. The mechanical property of the FG/BMIMCl composite material under compressive mode was superior compared with the GG and LBG/BMIMCl composite materials. Then, FG films compatibilized with polymeric ionic liquids (PILs) were also prepared by in situ radical polymerization of polymerizable ionic liquids, 1-(3-acryloyloxypropyl)-3-vinylimidazolium bromide and 1-methyl-3-vinylbenzylimidazolium chloride by AIBN in mixtures of FG with BMIMCl. The mechanical properties of the resulting films were affected by the FG/PIL ratios as well as the unit ratios in PILs.

Early and accurate diagnosis of invasive aspergillosis (IA) is one of themost critical steps needed to efficiently treat the infection and reduce the high mortality rates that can occur. We have previously found that the Aspergillus spp. secondary metabolite, bis(methylthio)gliotoxin (bmGT), can be detected in the serum from patients with possible/probable IA. Thus, it could be used as a diagnosis marker of the infection. However, there is no data available concerning the sensitivity, specificity and performance of bmGT to detect the infection. Here,we have performed a prospective study comparing bmGT detection with galactomannan (GM), the most frequently used and adopted approach for IA diagnosis, in 357 sera from 90 episodes of patients at risk of IA. Our results, involving 79 patients that finally met inclusion criteria, suggest that bmGT presents higher sensitivity and positive predictive value (PPV) than GM and similar specificity and negative predictive value (NPV). Importantly, the combination of GM and bmGT increased the PPV (100 %) and NPV (97.5 %) of the individual biomarkers, demonstrating its potential utility in empirical antifungal treatment guidance and withdrawal. These results indicate that bmGT could be a good biomarker candidate for IA diagnosis and, in combination with GM, could result in highly specific diagnosis of IA and management of patients at risk of infection.

Crude locust bean gum (CLBG) was purified and fractionated into two parts: the first was obtained by solubilization in water at 25°C (GM25) and the second consisted in a further extraction at 80°C on the residual impoverished fraction (GM80). The complete structural characterization has shown that GM80 possessed relatively longer chain lengths than GM25, a slightly lower degree of galactose substitution and a somewhat sharper galactosyl distribution in substituted and unsubstituted regions. A physical behavior analysis was carried out on solubilization kinetics, viscosity, viscoelasticity and formation of associated gels with xanthan or carrageenan. The average structure of GM80 generated larger intra-chain, inter-chain and inter-molecular interactions, resulting in the appearance of a stronger network. Small structural differences therefore generated very different physical behaviors. This study thus allowed to establish, in a precise and complete manner, fractionation-purification-structure-function relationships of galactomannans extracted from carob., info:eu-repo/semantics/published

The rheological behavior of galactomannans in concentrated solutions was examined by using dynamic viscoelasticity measurements. Concentrated solutions of three galactomannans, guar gum, tara gum, and locust bean gum were prepared with an ionic liquid 1-butyl-3-methylimidazolium chloride as the solvent. Each galactomannan solution showed angular frequency dependence curves of the storage modulus and the loss modulus which were characteristic of a solution of entangled polymer chains. The molecular weight between entanglements (Me) was obtained from the plateau modulus and the concentration dependence of Me showed Me in the molten state (Me, melt) to be 4.6 × 10[3], 3.2 × 10[3], and 2.7 × 10[3] for guar gum, tara gum, and locust bean gum, respectively. It was found that the material constant Me, melt depends on the mannose/galactose ratio of the galactomannans. The number of monosaccharide units between entanglements in the molten state for the galactomannans varied within the range found for other polysaccharides such as cellulose and agarose in ionic liquids, suggesting that all the galactomannans take a random-coil conformation in ionic liquid solutions.

Diagnosing chronic pulmonary aspergillosis (CPA) is complicated, and there are limited data available regarding the identification of galactomannan (GM) in clinical specimens to assist the detection of this infection. The purpose of this study was to evaluate the detection of GM in bronchoalveolar lavage fluid (BALF) and serum and to assess its utility for diagnosing CPA. We retrospectively reviewed the diagnostic and clinical characteristics of 144 patients, with and without CPA, in Nagasaki University Hospital, Japan, whose BAL and serum specimens were examined for the presence of GM. The Platelia Aspergillus enzyme immunoassay (PA EIA) was performed according to the manufacturer's instructions. The mean values of BALF GM antigen were 4.535 (range, 0.06214.120) and 0.430 (range, 0.0629.285) in CPA (18) and non-CPA (126) patients, respectively. The mean values of serum GM antigen were 1.557 (range, 0.2325.397) and 0.864 (range, 0.0288.956) in CPA and non-CPA patients, respectively. PA EIA of BALF is superior to the test with serum, with the optimal cut-off values for BALF and serum of 0.4 and 0.7, respectively. The sensitivity and specificity of PA EIA in BALF at a cut-off of 0.4 were 77.2% and 77.0%, respectively, whereas with serum at a cut-off of 0.7, they were 66.7% and 63.5%, respectively. GM testing using BALF showed reasonable sensitivity and specificity as compared to that using serum. Thus, assessing GM levels in BALF may enhance the accuracy of diagnosing CPA., Medical Mycology, 50(8), pp.811-817; 2012

Galactomannan (GM) is used to diagnose aspergillosis. We present a case of a hematopoietic stem cell transplantation recipient with fusariosis who received early antifungal treatment based on GM positivity. Additionally, 3 Fusarium isolates tested positive for GM. Fusarium is another mold containing GM. GM might be useful for diagnosing fusariosis in high-risk patients.

Suspenzní bujónovou metodou byl zkoumán pozitivní, případně negativní vliv galaktomananu, glukomananu, chitosanu a derivátů celulózy na probiotické bakterie Lactobacillus lactis a Enterococcus faecalis. Testované vzorky byly použity v koncentrační řadě 1, 2 a 3 hm %. U chitosanu byl prokázán inhibiční účinek ve všech použitých koncentracích. Glukomanan a hydroxyethylcelulóza působily po 3 dnech inkubace na snížení počtu buněk v menší míře, u Enterococcus faecalis došlo k inhibici růstu buněk pouze o 1 řád, počty Lactobacillus lactis nebyly ovlivněny vůbec. Mikrokrystalická celulóza a karboxymethylcelulóza měly na růst probiotických bakterií pozitivní účinek ve všech použitých koncentracích. Galaktomanan měl nejvýraznější pozitivní účinek při 2 % přídavku. Suspenzní bujónovou metodou a difuzní diskovou metodou byly zkoumány antimikrobiální účinky karboxymethylcelulózy a její kyselé formy v koncentrační řadě 2 a 3 hm % pro KMC a 1, 2 a 3 hm % pro CMC. Dále byly zkoumány účinky při interakci obou derivátů celulózy v příslušné koncentrační řadě s 2 a 5 % roztokem Betadinu. Karboxymethylcelulóza neměla inhibiční účinky v žádné použité koncentraci, zatímco její kyselá forma inhibovala bakterii Pseudomonas aeruginosa ve všech koncentracích a bakterii Staphylococcus aureus pouze v 2 a 3 % roztocích. Karboxymethylcelulóza a její kyselá forma v interakci s Betadinem účinně inhibovaly obě používané bakterie ve všech zkoumaných koncentracích., Positive or negative effects of galactomannan, glucomannan, chitosan and derivates of celulose were studied with suspenzion broth method on probiotic bacteria Lactobacillus lactis and Enterococcus faecalis. The samples were tested in the 1, 2 and 3 % concentrations. In all used concentrations chitosan has shown chitosanu inhibitory effects. After 3 days of incubation it was observed the reduction of numbers by glucomannan and hydroxyethylcellulose in all used concetrations too. There were achieved positive effects were with microcrystalline celulose and carboxymethylcellulose. With 2 % addition of galactomannan had the most positive effect. Antimicrobial effects of carboxymethylcellulose and its acid form were investigated in concentration range of 1, 2 and 3 %. Antimicrobial effects of these two polysaccharides were investigated in the interaction with the concentration range of 2 and 5 % Betadin. All used concentrations of carboxymethylcellulose did not caused an inhibitory effects. The acid form of carboxymethylcellulose inhibited Pseudomonas aeruginosa at all levels and Staphylococcus aureus in only 2 and 3 % solutions. In all used concentrations of carboxymethylcellulose and its acid form in the interaction with Betadin inhibited both the bacteria., Katedra analytické chemie, Dokončená práce s úspěšnou obhajobou

At present, voriconazole (VOR) is the drug of first choice for treating invasive pulmonary aspergillosis (IPA). However, particularly in advanced stages of disease and in the severely immunocompromised host, the mortality remains substantial. The combination of VOR with an echinocandin may improve the therapeutic outcome. We investigate here whether combining VOR and anidulafungin (ANI) in advanced IPA in transiently neutropenic rats results in a higher therapeutic efficacy. Since VOR is metabolized more rapidly in rodents than in humans, dosage adjustment for VOR is necessary to obtain an area under the plasma concentrationtime curve (AUC) in rodents that is equivalent to that of humans. In this study, the pharmacokinetics of VOR and ANI in rats were elucidated, and dosage schedules were applied that produced AUCs similar to those of humans. The developed dose schedules were well tolerated by the rats, without effects on renal and hepatic functions. VOR showed excellent efficacy in early IPA (100% rat survival). In advanced IPA, VOR was less efficacious (50% rat survival), whereas a significant decrease in galactomannan concentrations in lungs and sera was found in surviving rats. ANI administered in advanced IPA resulted in 22% rat survival, and the serum concentrations of fungal galactomannan were slightly but not significantly decreased. The addition of ANI to VOR did not result in significantly increased therapeutic effic

Serology is currently used for the diagnosis of canine sino-nasal aspergillosis (SNA). However, the accuracy of serological testing using commercially available, standardized purified antigen preparations of Aspergillus (CAPurAspAg) has only been poorly documented. The aim of the present study was to assess the diagnostic value of an agar-gel double immunodiffusion (AGDD) test and an anti-Aspergillus IgG ELISA, using CAPurAspAg and the commercially available Platelia™ test for the detection of serum galactomannan. Sera from 17 dogs with SNA, 18 dogs with a nasal tumour (NT), 11 dogs with lymphoplasmacytic rhinitis (LPR) and 33 control dogs were tested with the 3 methods. AGDD result was positive in 76.5% of dogs with SNA, whereas all sera from dogs with non-fungal nasal disease and control dogs were negative. A positive IgG ELISA result was obtained in 88% of dogs with SNA and in 18% of dogs with LPR. All patients with NT and control dogs had a negative IgG ELISA result. The Platelia™ test was positive in 24% of dogs with SNA, 11% of dogs with NT, 9% of dogs with LPR and 24% of control dogs. The results of this study suggest that (1) the detection of serum Aspergillus-specific antibodies with AGDD or ELISA, using CAPurAspAg, provides excellent specificity and good sensitivity, (2) the specificity is higher for AGDD (100%) than for ELISA (96.8%) while sensitivity is higher for ELISA (88.2%) than for AGDD (76.5%) and (3) serum galactomannan quantification with the Platelia™ test is unreliable for the diagnosis of canine SNA. © 2008 Elsevier B.V. All rights reserved., SCOPUS: ar.j, info:eu-repo/semantics/published