Optimization of solid-state fermentation for enhanced production of pectinolytic complex by Aspergillus tubingensis FAT43 and its application in fruit juice processing.

The main goal of this study was to examine the efficiency of a newly isolated fungus from quince, Aspergillus tubingensis FAT43, to produce the pectinolytic complex using agricultural and industrial waste as the substrate for solid state fermentation. Sugar beet pulp was the most effective substrate inducer of pectinolytic complex synthesis out of all the waste residues examined. For endo-pectinolytic and total pectinolytic activity, respectively, statistical optimization using Placked-Burman Design and Optimal (Custom) Design increased production by 2.22 and 2.15-fold, respectively. Liquification, clarification, and an increase in the amount of reducing sugar in fruit juices (apple, banana, apricot, orange, and quince) processed with pectinolytic complex were identified. Enzymatic pre-treatment considerably increases yield (14%-22%) and clarification (90%). After enzymatic treatment, the best liquefaction was observed in orange juice, whereas the best clarification was obtained in apricot juice. Additionally, the pectinolytic treatment of apricot juice resulted in the highest increase in reducing sugar concentration (11%) compared to all other enzymatically treated juices. Optimizing the production of a highly active pectinolytic complex and its efficient utilization in the processing of fruit juices, including the generation of an increasing amount of waste, are the significant outcomes of this research.

Sources and a Health Risk Assessment of Potentially Toxic Elements in Dust at Children's Playgrounds with Artificial Surfaces: A Case Study in Belgrade.

The focus of this research on children's playgrounds with artificial surfaces aimed to establish levels of potentially toxic elements (PTEs) in dust, their origin, and impact on children at 15 playgrounds: 9 on school grounds and 6 on day nurseries in Belgrade (Serbia). Soil samples were taken from the immediate vicinity of the playgrounds to establish the origin of PTEs in the dust samples. Soil analyses revealed the lithogenic origin of Co, Cr, Ni, Fe, Mn, As, Cd, Cu, Pb, and Zn and the anthropogenic origin of As, Cd, Cu, Pb, and Zn. However, in the dust samples, the origin of the elements was different with As, Co, Fe, and Mn originating from the surrounding soil; Cr and Ni levels affected by both atmospheric deposition and the surrounding soil; Cd, Pb, and Zn concentrations impacted by atmospheric deposition; and Cu levels affected by factors of a local character. No noncancer risk was found for any of the individual elements investigated, nor for any of the playgrounds being studied, while a minimal cancer risk was found from As with values greater than 1E-6 at almost all the sites. Based on the results obtained for the spatial distribution of individual PTE levels, it was determined that the surrounding soil and atmospheric deposition have an almost equal impact on noncancer risk values.

Evaluation of Salix alba, Juglans regia and Populus nigra as biomonitors of PTEs in the riparian soils of the Sava River.

A large number of human activities result in the release of potentially toxic elements (PTEs) into the environment, which could lead to the degradation of riparian areas. This study aimed to evaluate the potential of Salix alba, Juglans regia and Populus nigra for the biomonitoring of PTEs in the riparian soils of the Sava River. Levels of seven PTEs (As, Cd, Cr, Cu, Ni, Pb and Zn) were measured in the soils, roots and leaves of plants at selected sampling sites and evaluated according to bioaccumulation and translocation factors. The obtained results showed that in riparian soils, As, Cr, Cu, Ni and Zn were at levels considered to be critical for plants. The levels of As, Cd, Cr, Ni and Zn measured in roots of Salix alba and As, Cr, Ni and Zn in its leaves were toxic for plant tissue. Toxic levels of Cr were also measured in the roots of Juglans regia and As in its leaves, as well as As and Cr in the roots of Populus nigra, and Zn in its leaves. Bioconcentration and translocation factors showed that S. alba and P. nigra have potential for the phytoextraction of Zn and Cd, while J. regia has potential for the phytoextraction of As. In terms of phytostabilization potential, S. alba proved to be good for the phytostabilization of Cd and Cu, and J. regia for the phytostabilization of Cr, As, Ni and Pb, while P. nigra showed potential for the phytostabilization of Cr, Ni, Pb and Cu.

Fractionation, Mobility, and Contamination Assessment of Potentially Toxic Metals in Urban Soils in Four Industrial Serbian Cities.

The main soil properties, concentrations of selected elements (As, Cd, Cr, Cu, Fe, Mn, Ni, Pb, and Zn), and the chemical speciation of each element were determined in urban soil samples taken from urban parks in four Serbian cities (Belgrade, Pancevo, Obrenovac, and Smederevo) exposed to different sources of pollution. Pollution indices (PI, PIN) and factors (MF, ICF, GCF) also were evaluated. The study revealed As and Cd concentrations below the detection limit, whereas the content of Cr, Cu, Fe, Mn, Ni, Pb, and Zn at some sites exceeded the limits established by local regulations, as well as the background values, which may represent an environmental threat. Sequential extraction results show that Fe, Cr, Cu, and Ni were predominantly in the residual fraction at most sites; however, Ni from Pancevo and Smederevo also was bound to the reducible fraction. The presence of Pb at all sites and Zn in Smederevo and Belgrade was mainly associated with the reducible and residual fractions. The highest Mn content was found in the reducible fraction, followed by the acid soluble/exchangeable and residual fractions. Based on the obtained indices and factors, the overall soil status at the selected sampling sites was found to range from the warning limit to slightly polluted, whereby Smederevo had the highest risk, and Pancevo and the control site the lowest risk of contamination by toxic metals.

Plant growth-promoting bacteria elevate the nutritional and functional properties of black cumin and flaxseed fixed oil.

BACKGROUND: In order to study the influence of plant growth-promoting bacteria (PGPB) belonging to Streptomyces sp., Paenibacillus sp., and Hymenobacter sp. on fixed oil content of flaxseed and black cumin, 2-year field experiments were conducted. PGPB was applied during seedtime of plants. The extraction of oil from seeds was performed using supercritical CO2 . RESULTS: The addition of PGPB significantly increases the content of C18:1 (from 16.06 ± 0.03% to 16.97 ± 0.03%) and C18:3 (from 42.97 ± 0.2% to 45.42 ± 0.5%) in flaxseed oil and C18:2 (from 52.68 ± 0.50% to 57.11 ± 0.40%) and C20:2 (from 4.34 ± 0.02% to 4.54 ± 0.03%) in black cumin seed oil. The contents of total polyphenols, flavonoids, and carotenoids, as well as antioxidant activity measured by ferric-reducing ability of plasma assay, were found to be greater in the oil from the seeds of plants treated with the PGPB, compared with the respective non-treated samples. CONCLUSION: The use of PGPB enhances plant nutritive properties; these represent a great source for obtaining valuable functional food ingredients. © 2017 Society of Chemical Industry.

Allergen relative abundance in several wheat varieties as revealed via a targeted quantitative approach using MS.

Food allergy has become a major health issue in developed countries, therefore there is an urgent need to develop analytical methods able to detect and quantify with a good sensitivity and reliability some specific allergens in complex food matrices. In this paper, we present a targeted MS/MS approach to compare the relative abundance of the major recognized wheat allergens in the salt-soluble (albumin/globulin) fraction of wheat grains. Twelve allergens were quantified in seven wheat varieties, selected from three Triticum species: T. aestivum (bread wheat), T. durum (durum wheat), and T. monococcum. The allergens were monitored from one or two proteotypic peptides and their relative abundance was deduced from the intensity of one fragment measured in MS/MS. Whereas the abundance of some of the targeted allergens was quite stable across the genotypes, others like alpha-amylase inhibitors showed clear differences according to the wheat species, in accordance with the results of earlier functional studies. This study enriches the scarce knowledge available on allergens content in wheat genotypes, and brings new perspectives for food safety and plant breeding.

Cell wall proteomic of Brachypodium distachyon grains: A focus on cell wall remodeling proteins.

Cell walls play key roles during plant development. Following their deposition into the cell wall, polysaccharides are continually remodeled according to the growth stage and stress environment to accommodate cell growth and differentiation. To date, little is known concerning the enzymes involved in cell wall remodeling, especially in gramineous and particularly in the grain during development. Here, we investigated the cell wall proteome of the grain of Brachypodium distachyon. This plant is a suitable model for temperate cereal crops. Among the 601 proteins identified, 299 were predicted to be secreted. These proteins were distributed into eight functional classes; the class of proteins that act on carbohydrates was the most highly represented. Among these proteins, numerous glycoside hydrolases were found. Expansins and peroxidases, which are assumed to be involved in cell wall polysaccharide remodeling, were also identified. Approximately half of the proteins identified in this study were newly discovered in grain and were not identified in the previous proteome analysis conducted using the culms and leaves of B. distachyon. Therefore, the data obtained from all organs of B. distachyon infer a global cell wall proteome consisting of 460 proteins. At present, this is the most extensive cell wall proteome of a monocot species.

Unveiling novel insights into Bacillus velezensis 16B pectin lyase for improved fruit juice processing.

Microbial pectinolytic enzymes are important in various industries, particularly food processing. This study focuses on uncovering insights into a novel pectin lyase, BvPelB, from Bacillus velezensis 16B, with the aim of enhancing fruit juice processing. The study examines the structural and functional characteristics of pectinolytic enzyme, underscoring the critical nature of substrate specificity and enzymatic reaction mechanisms. BvPelB was successfully expressed and purified, exhibiting robust activity under alkaline conditions and thermal stability. Structural analysis revealed similarities with other pectin lyases, despite limited sequence identity. Biochemical characterization showed BvPelB's preference for highly methylated pectins and its endo-acting mode of cleavage. Treatment with BvPelB significantly increased juice yield and clarity without generating excessive methanol, making it a promising candidate for fruit juice processing. Overall, this study provides valuable insights into the enzymatic properties of BvPelB and its potential industrial applications in improving fruit juice processing efficiency and quality.

Improvement of fruit juice quality: novel endo-polygalacturonase II from Aspergillus tubingensis FAT 43 for enhanced liquefaction, clarification, and antioxidant potential.

This study focuses on the isolation, purification, and characterisation of endo-polygalacturonase II from Aspergillus tubingensis FAT43, particularly emphasising its potential applications in the fruit juice industry. A comprehensive screening test revealed the temporal dynamics of endo-polygalacturonase production during a 96-hour fermentation process. The purification process, involving ammonium sulfate and ethanol precipitation followed by ion-exchange chromatography, resulted in a 3.3-fold purification of PG II with a yield of 16% and a specific activity of 6001.67 U mg-1. Molecular analysis confirmed the identity of PG II, its gene (pgaII), and a high degree of sequence identity with Aspergillus tubingensis in the SWISS-PROT database. The optimal pH for PG II activity was 3.5-4.5, with robust stability across a broad pH spectrum (3-7). The enzyme exhibited optimal temperature activity at 45 °C, with a retention of 90% activity at 50 °C. The calculated activation energy for PG II was 62.1 kJ mol-1, indicating good stability. Inactivation kinetics revealed a half-life of 13.7 h at 40 °C, 5.4 h at 50 °C, and 0.85 h at 60 °C, with an activation energy of denaturation of 32.8 kJ mol-1. Compared to literature-reported PGs, PG II from A. tubingensis FAT43 demonstrated superior thermal stability. Hydrolysis experiments on different pectins revealed the highest specificity for non-methylated substrates (polygalacturonic acid). In fruit juice processing, PG II significantly increased juice yield and clarity, with the highest impact observed in strawberry juice. Antioxidant activity assays indicated enhanced antioxidant potential in enzyme-treated juices, especially strawberry, quince, and apple juices. The study highlights PG II's potential as an industrially valuable enzyme for fruit juice processing, offering improved thermostability and versatility across various fruit types.

Sex determination in 58 bird species and evaluation of CHD gene as a universal molecular marker in bird sexing.

The aim of this research was to test the CHD gene (Chromo Helicase DNA-binding gene) as a universal molecular marker for sexing birds of relatively distant species. The CHD gene corresponds to the aim because of its high degree of conservation and different lengths in Z and W chromosomes due to different intron sizes. DNA was isolated from feathers and the amplification of the CHD gene was performed with the following sets of polymerase chain reaction (PCR) primers: 2550F/2718R and P2/P8. Sex determination was attempted in 284 samples of 58 bird species. It was successful in 50 bird species; in 16 of those (Alopochen aegyptiacus, Ara severus, Aratinga acuticaudata, Bucorvus leadbeateri, Cereopsis novaehollandiae, Columba arquatrix, Corvus corax, C. frugilegus, Cyanoliseus patagonus, Guttera plumifera, Lamprotornis superbus, Milvus milvus, Neophron percnopterus, Ocyphaps lophotes, Podiceps cristatus, and Poicephalus senegalus), it was carried out for the first time using molecular markers and PCR. It is reasonable to assume that extensive research is necessary to define the CHD gene as a universal molecular marker for successful sex determination in all bird species (with exception of ratites). The results of this study may largely contribute to the aim.

BrainLine: An Open Pipeline for Connectivity Analysis of Heterogeneous Whole-Brain Fluorescence Volumes.

Whole-brain fluorescence images require several stages of computational processing to fully reveal the neuron morphology and connectivity information they contain. However, these computational tools are rarely part of an integrated pipeline. Here we present BrainLine, an open-source pipeline that interfaces with existing software to provide registration, axon segmentation, soma detection, visualization and analysis of results. By implementing a feedback based training paradigm with BrainLine, we were able to use a single learning algorithm to accurately process a diverse set of whole-brain images generated by light-sheet microscopy. BrainLine is available as part of our Python package brainlit: http://brainlit.neurodata.io/ .

Different K+ channels are involved in relaxation of arterial and venous graft induced by nicorandil.

The drug nicorandil is a vasodilator approved for the treatment of angina. In addition to its well-known effect on the opening of ATP-sensitive K (KATP) channels, nicorandil-induced vasorelaxation also involves the opening of Ca-activated K channels. The aim of this study was to investigate the effects of nicorandil on the isolated human internal mammary artery (HIMA) and the human saphenous vein (HSV) and to define the contribution of different K channel subtypes in the nicorandil action on these arterial and venous grafts. Our results show that nicorandil induced a concentration-dependent relaxation of HSV and HIMA rings precontracted by phenylephrine. Glibenclamide, a selective KATP channels inhibitor, partially inhibited the response to nicorandil in both HSV and HIMA. Iberiotoxin, a most selective blocker of large-conductance Ca-activated K (BKCa) channels, partly antagonized relaxation of HIMA. A nonselective blocker of voltage-gated K channels, 4-aminopyridine caused partial inhibition of the nicorandil-induced relaxation of HSV but did not antagonize relaxation of HIMA induced by nicorandil. Margatoxin, a potent inhibitor of KV1.3 channels, did not abolish the effect of nicorandil on HSV and HIMA. Our results showed that nicorandil induced strong endothelium-independent relaxation of HSV and HIMA contracted by phenylephrine. It seems that KATP and 4-aminopyridine-sensitive K channels located in the smooth muscle of HSV mediated relaxation induced by nicorandil. In addition, KATP and BKCa channels are probably involved in the nicorandil action on HIMA.

Development and validation of an HPLC method for determination of ziprasidone and its impurities in pharmaceutical dosage forms.

Ziprasidone is known as a novel "atypical" or "second-generation" antipsychotic drug. A sensitive and reproducible method was developed and validated for determination of ziprasidone and its major impurities, which are significantly different in polarity. The separation is performed on a Waters Spherisorb octadecylsilyl 1 column (5.0 microm particle size, 250 x 4.6 mm id) using a gradient with mobile phase A [buffer-acetonitrile (80+20, v/v)] and mobile phase B [buffer-acetonitrile (10+90, v/v)] at a working temperature of 25 degrees C. The buffer was 0.05 M KH2PO4 solution with an addition of 10 mL triethylamine/L solution, adjusted to pH 2.5 with orthophosphoric acid. The flow rate was 1.5 mL/min, and the eluate was monitored at 250 nm using a diode array detector. Optimization of the experimental conditions was performed using partial least squares regression, for which four factors were selected for optimization: buffer concentration, buffer pH, triethylamine concentration, and temperature. The proposed validated method is convenient and reliable for the assay and purity control in both raw materials and dosage forms.

Risk factors associated with carotid artery puncture following landmark-guided internal jugular vein cannulation attempts.

OBJECTIVE: The relationship between certain risk factors and carotid artery puncture (CAP) as an early mechanical complication following internal jugular vein cannulation attempts (IJVCAs) was evaluated. METHODS: In a retrospective 1-year observational single-center study, 86 IJVCAs conducted in the operating room by 4 competent anesthesiologists were evaluated. Age, gender, puncture side, number of cannulation attempts, circumstances of the procedure and incidence of CAP were obtained from medical records. RESULTS: Of the 86 IJVCAs performed in patients aged 18-75 years, CAP occurred in 8 (9.3%): 5 (5.8%) in patients >65 years and 3 (3.5%) in patients <65 years of age. CAP was not associated with patient's age (p = 0.11) and gender (p = 0.76). Multiple cannulation attempts (OR = 26.25; 95% CI = 4.52-152.51; p < 0.001) and placement of CVC under emergency conditions (OR = 14.84; 95% CI = 1.73-127.22; p = 0.014) increased the risk for CAP significantly. Also, the risk for CAP was higher when IJVCAs were performed before induction of general anesthesia (OR = 15.75; 95% CI = 1.83-135.1; p = 0.019). CAP was more likely to happen during left-sided than right-sided IJVCA (OR = 5.98; 95% CI = 1.29-27.59; p = 0.022). In addition, left-sided attempts considerably increased the risk for multiple cannulation attempts (OR = 2.782; 95% CI = 1.342-3.965; p < 0.01). Also, manifold cannulation attempts were more frequent if the IJVCA was performed before induction of anesthesia (OR = 4.219; CI = 1.579-11.271; p = 0.004). CONCLUSIONS: Our results strongly suggest that left-sided, multiple IJVCAs, performed under emergency conditions in conscious patients in the operating room, represent considerable risks for possible CAP.

Chemical Fractionation, Environmental, and Human Health Risk Assessment of Potentially Toxic Elements in Soil of Industrialised Urban Areas in Serbia.

The primary focus of this research was the chemical fractionation of potentially toxic elements (PTEs) and their presence in several industrialised cities in Serbia. Furthermore, their origin, contamination levels, and environmental and human health risks were assessed. The results indicated that the examined soils were characterised by slightly higher Cu, Ni, Pb, and Zn levels than those set by European and national regulations. These elevated Cu, Pb, and Zn concentrations were caused by intensive traffic and proximity to industry, whereas the higher Ni levels were a result of the specific geological substrate of the soil in the study area. The environmental risk was found to be low and there was no enrichment/contamination of the soil with these elements, except in the case of Pb, for which moderate to significant enrichment was found. Lead also poses a potential non-carcinogenic risk to children through ingestion and requires special attention due to the fact that a significant proportion of this element was present in the tested soil samples in a potentially available form. Analysis of the health risks showed that children are more at risk than adults from contaminants and that ingestion is the riskiest exposure route. The carcinogenic risk was within the acceptable limits.

Fractionation of Potentially Toxic Elements (PTEs) in Urban Soils from Salzburg, Thessaloniki and Belgrade: An Insight into Source Identification and Human Health Risk Assessment.

Concentrations of potentially toxic elements (PTEs) (Al, As, Cd, Cr, Cu, Ni, Pb, and Zn) were measured in topsoil samples collected from parks in the cities of Salzburg (Austria), Thessaloniki (Greece), and Belgrade (Serbia) in order to assess the distribution of PTEs in the urban environment, discriminate natural (lithogenic) and anthropogenic contributions, identify possible sources of pollution, and compare levels of pollution between the cities. An assessment of the health risks caused by exposure to PTEs through different pathways was also conducted. The study revealed that, with the exception of Pb in Salzburg, levels of PTEs in the soils in polluted urban parks were higher than in unpolluted ones, but still lower than those recorded in other European soils. Results of sequential analyses showed that Al, Cr, and Ni were found in residual phases, proving their predominantly lithogenic origin and their low mobility. In contrast, the influence of anthropogenic factors on Cu, Pb, and Zn was evident. Site-dependent variations showed that the highest concentrations of As, Cu, Pb, and Zn of anthropogenic origin were recorded in Salzburg, while the highest levels of Al, Cr, and Ni of lithogenic origin were recorded in Belgrade and Thessaloniki, which reflects the specificity of the geological substrates. Results obtained for the health risk assessment showed that no human health risk was found for either children or adults.

Influence of breed on selected quality parameters of fresh goat meat.

The potential of goats to produce a high-quality meat is mainly reflected in their healthy fats, low calorie intramuscular fats, saturated fats, and, especially, their high ratios of unsaturated (UFA) and saturated (SFA) fatty acids, as well as hypocholesterolemic and hypercholesterolemic fatty acids. The aim of this study was to collect and compare meat quality parameters for domestic Balkan, Alpine and Saanen goats of the same age. Samples for all tests were taken from musculus gluteus superficialis. Chemical composition, pH value, fatty acid composition, content of volatile compounds, color and overall sensory quality (appearance, texture and smell) were determined. In chemical composition, moisture, fat, protein and ash varied significantly between each of the examined groups as opposed to pH values. Furthermore, among all the examined groups a significant difference was found for fatty acids and volatile compounds. Determined ratio of polyunsaturated fatty acids (PUFAs) to SFAs was 0.089, 0.085 and 0.071 for Balkan, Alpine and Saanen goat meats, respectively. Regarding that ratio, Saanen goat meat had the most favorable characteristics. Saanen goat meat showed the highest nutritional value. On the other hand, Balkan goat meat had the lowest intramuscular fat content. Measurements of the meat color from all three groups, as well as overall acceptability, showed significant differences between breeds. Obtained results point to the impact of breed on chemical composition and fatty acid profile of goat meat.

A surgical reentry study on the influence of platelet-rich plasma in enhancing the regenerative effects of bovine porous bone mineral and guided tissue regeneration in the treatment of intrabony defects in humans.

BACKGROUND: The purpose of this study was to evaluate the additional benefits provided by the incorporation of platelet-rich plasma (PRP) into a regenerative protocol consisting of bovine porous bone mineral (BPBM) and guided tissue regeneration (GTR) in the treatment of intrabony defects in humans. METHODS: Twenty-three paired intrabony defects were surgically treated using a split-mouth design. Defects were treated with BPBM/GTR/PRP (experimental group) or with BPBM/GTR (control group). The clinical parameters evaluated included changes in probing depth, clinical attachment level, and defect fill as revealed by reentry surgeries at 6 months. RESULTS: Preoperative probing depths, attachment levels, and transoperative bone measurements were similar for the two groups. Post-surgical measurements taken at 6 months revealed that both treatment modalities resulted in a significant decrease in probing depth, gain in clinical attachment, and bone fill of the defects compared to baseline. Postoperative differences observed between the two groups were 0.72 +/- 0.36 mm at buccal sites and 0.90 +/- 0.32 mm at lingual sites for probing depth, 0.82 +/- 0.41 mm at buccal sites and 0.78 +/- 0.38 at lingual sites for gain in clinical attachment, and 0.85 +/- 0.36 mm at buccal sites and 0.94 +/- 0.42 mm at lingual sites for defect fill, all favoring the experimental sites. However, none of the differences were statistically significant. CONCLUSION: Within the limitations related to using a small sample size, PRP did not significantly augment the effects of BPBM and GTR in promoting the clinical resolution of intrabony defects.

Customizing the spent coffee for Trichoderma reesei cellulase immobilization by modification with activating agents.

This study has explored the possibility to reuse the waste, spent coffee material for the cellulase enzyme immobilization. By the coffee surface modification with different activating agents, it was attempted to develop the convenient method for creation of a capable porous carrier for this purpose. Among the most common activating agents, glutaraldehyde, chlorine dioxide and hydrogen peroxide provided the most acceptable choice for the coffee surface modification. The changes that occurred on the coffee surface due to agents' treatment exposure were recorded by using of the FTIR spectra and SEM micrographs. The highest immobilization yield (55%) and immobilization efficiency (45%) were attained during 30min of the treatment time, by employing of 30% chlorine dioxide aqueous solution within 6mL/g activator/carrier ratio. The kinetic process was found to be predicted by the pseudo-second-order model. The cellulase immobilization onto the coffee surface provides an excellent base for increasing the enzyme availability to the substrate and enhancing the enzyme productivity, by offering the new perspectives to the industrial sector.