Stabilization of Black Locust Flower Extract via Encapsulation Using Alginate and Alginate-Chitosan Microparticles.

Black locust flower extract contains various polyphenols and their glucosides contribute to the potential health benefits. After intake of these bioactive compounds and passage through the gastrointestinal tract, their degradation can occur and lead to a loss of biological activity. To overcome this problem, the bioactive compounds should be protected from environmental conditions. This study aimed to encapsulate the black flower extract in the microparticles based on biodegradable polysaccharides, alginate, and chitosan. In the extract, the total antioxidant content was found to be 3.18 ± 0.01 g gallic acid equivalent per 100 g of dry weight. Also, the presence of lipids (16), phenolics (27), organic acids (4), L-aspartic acid derivative, questinol, gibberellic acid, sterol, and saponins (2) was confirmed using the UHPLC-ESI-MS analysis. In vitro assays showed that the extract has weak anti-α-glucosidase activity and moderate antioxidant and cytotoxic activity against the HeLa cell line. The extrusion method with secondary air flow enabled the preparation of microparticles (about 270 µm) encapsulated with extract. An encapsulation efficiency of over 92% was achieved in the alginate and alginate-chitosan microparticles. The swelling study confirmed a lower permeability of alginate-chitosan microparticles compared with alginate microparticles. For both types of microparticles, the release profile of antioxidants in the simulated gastrointestinal fluids at 37 °C followed the Korsmeyer-Peppas model. A lower diffusion coefficient than 0.5 indicated the simple Fick diffusion of antioxidants. The alginate-chitosan microparticles enabled a more sustained release of antioxidants from extract compared to the alginate microparticles. The obtained results indicated an improvement in the antioxidant activity of bioactive compounds from the extract and their protection from degradation in the simulated gastric conditions via encapsulation in the polymer matrixes. Alginate-chitosan showed slightly slower cumulative antioxidant release from microparticles and better antioxidant activity of the extract compared to the alginate system. According to these results, alginate-chitosan microparticles are more suitable for further application in the encapsulation of black locust flower extract. Also, the proposed polymer matrix as a drug delivery system is safe for human use due to its biodegradability and non-toxicity.

Preparation and Characterization of Alginate Hydrogels with High Water-Retaining Capacity.

Hydrogels are very attractive materials due to their multifunctional properties. Many natural polymers, such as polysaccharides, are used for the preparation of hydrogels. The most important and commonly used polysaccharide is alginate because of its biodegradability, biocompatibility, and non-toxicity. Since the properties of alginate hydrogel and its application depend on numerous factors, this study aimed to optimize the gel composition to enable the growth of inoculated cyanobacterial crusts for suppressing the desertification process. The influence of alginate concentration (0.1-2.9%, m/v) and CaCl2 concentration (0.4-4.6%, m/v) on the water-retaining capacity was analyzed using the response surface methodology. According to the design matrix, 13 formulations of different compositions were prepared. The water-retaining capacity was defined as the system response maximized in optimization studies. The optimal composition of hydrogel with a water-retaining capacity of about 76% was obtained using 2.7% (m/v) alginate solution and 0.9% (m/v) CaCl2 solution. Fourier transform infrared spectroscopy was used for the structural characterization of the prepared hydrogels, while the water content and swelling ratio of hydrogels were determined using gravimetric methods. It was concluded that alginate and CaCl2 concentrations play the most important role regarding the gelation time, homogeneity, water content, and swelling ratio of the hydrogel.

Sustainable Dyeing and Functionalization of Different Fibers Using Orange Peel Extract's Antioxidants.

A diluted ethanol orange peel extract was used for sustainable dyeing and functionalization of different fabrics. The extract analysis was performed using UPLC-ESI-MS/MS; its total flavonoid (0.67 g RE/100 g d.w.) and antioxidant (2.81 g GAE/100 g d.w.) contents and antioxidant activity (IC50 of 65.5 µg/mL) were also determined. The extract dyeing performance at various dyebath pH values was evaluated using multifiber fabric. Among six fabrics, extract possessed the ability for dyeing wool, polyamide, and cellulose acetate (at pH 4.5), which color strength (K/S) values increased after washing (9.7-19.8 vs. 11.6-23.2). Extract:water ratio of 20:35 (v/v) was found to be sufficient for achieving satisfactory K/S values (i.e., 20.17, 12.56, and 10.38 for wool, polyamide, and cellulose acetate, respectively) that were slightly changed after washing. The optimal dyeing temperatures for wool, polyamide, and cellulose acetate are 55, 35, and 25 °C, while the equilibrium dye exhaustion at those temperatures was achieved after 45, 120, and 90 min, respectively. The color coordinate measurements revealed that wool and polyamide fabrics are yellower than cellulose acetate, while, compared to polyamide and cellulose acetate, wool is redder. Possible interactions between selected fabrics and extract compounds are suggested. All fabrics possessed excellent antioxidant activity (88.6-99.6%) both before and after washing. Cellulose acetate provided maximum bacterial reduction (99.99%) for Escherichia coli, and Staphylococcus aureus, which in the case of Staphylococcus aureus remained unchanged after washing. Orange peel extract could be used for simultaneous dyeing and functionalization of wool and polyamide (excellent antioxidant activity) and cellulose acetate (excellent antioxidant and antibacterial activity) fabrics.

Optimization of Ultrasound-Assisted Extraction and Encapsulation of Antioxidants from Orange Peels in Alginate-Chitosan Microparticles.

The recovery of bioactive compounds from waste and modification of their properties by encapsulation are the main challenges today. In this study, the ultrasound-assisted extraction of antioxidants from orange peels was optimized using a central composite design. Ethanol (50%, v/v) was the solvent of choice for their extraction. The obtained total antioxidant contents were fitted using the second-order polynomial equation. The optimal conditions were the extraction time of 30 min, temperature of 60 °C, and the liquid-to-solid ratio of 15 mL/g. After that, the optimal extract was encapsulated in alginate-chitosan beads to modify the release of antioxidants under gastrointestinal tract conditions. The average size of beads was 252 µm, while the encapsulation efficiency was 89.2%. The results of the FTIR analysis indicated that there are no interactions between compounds of the extract and alginate-chitosan. In vitro release studies showed an initial rapid and then slow release of antioxidants. This release followed the simple Fickian diffusion. The encapsulation of orange peel extract provided improvement in the delivery of antioxidants after gastrointestinal digestion. The obtained encapsulated beads can be applied as the natural active ingredient of food, cosmetics, and pharmaceutical products.

Ultrasound-Assisted Extraction of Carotenoids from Orange Peel Using Olive Oil and Its Encapsulation in Ca-Alginate Beads.

The paper was aimed at developing an ultrasound-assisted extraction of carotenoids from orange peel using olive oil as a solvent. A central composite design was used to define the optimal conditions for their extraction. Under the optimal conditions (extraction time of 35 min, extraction temperature of 42 ℃, and a liquid-to-solid ratio of 15 mL/g), the experimental and predicted values of carotenoid content were 1.85 and 1.83 mg/100 g dry weight, respectively. The agreement of these values indicated the adequacy of the proposed regression model. The extraction temperature only had a negative influence on carotenoid content. The impact of extraction parameters on the carotenoid content was decreased according to the following order: extraction time, liquid-to-solid ratio, and extraction temperature. Ca-alginate beads were prepared using the extrusion process to increase the stability and protect the antioxidant activity of olive oil enriched with carotenoids. The encapsulation efficiency and particle mean diameter were 89.5% and 0.78 mm, respectively. The presence of oil extract in Ca-alginate beads was confirmed by Fourier-transform infrared spectroscopy. The antioxidant activity of the oil enriched with carotenoids before and after encapsulation in the alginate beads was determined according to the DPPH assay.

Optimization of ultrasound-assisted extraction of polyphenols from wheatgrass (Triticum aestivum L.).

Conventional extraction techniques require high consumption of available resources and thus are ineffective and expensive, especially at an industrial scale. The aim of the study was to optimize the ultrasound-assisted extraction of polyphenols from fresh wheatgrass (Triticum aestivum L.). The effects of different extraction techniques and solvents were investigated on the yield of extractive substances and antioxidant activity. The ultrasound-assisted extraction technique and ethanol gave the highest yield of extractive substances so that they were used in the optimization studies. The central composite design was employed to find the optimal levels of ethanol concentration, extraction temperature, and extraction time. The total phenolic content was varied in the range of 10.50-15.50 grams of gallic acid equivalents per 100 g of dry weight of plant material (g GAE 100 g-1 dw). The optimal conditions for ultrasound-assisted extraction were: (1) 56% (v/v) ethanol, (2) temperature of 59 °C, and (3) extraction time of 28 min. The results of ANOVA indicated that the highest impact had the extraction temperature on the total phenolic content. The toxic solvents were not used in the developed extraction procedure. The consumption of energy and raw plant material is estimated to be lower by at least 10% compared to conventional techniques.

Physico-Chemical Properties and Oxidative Stability of Fixed Oil from Plum Seeds (Prunus domestica Linn.).

Storage of a great amount of plum kernel waste becomes a challenge for food industry. In this work, the plum seed was used as a source of fixed oil that can be an ingredient of commercial products. Soxhlet extraction was carried out using the different solvents, such as n-hexane, n-heptane, ethyl acetate, acetone, or chloroform:methanol mixture (2:1 v/v). The highest yield of oil (about 30%) was obtained using n-heptane and n-hexane, while the lowest yield was obtained using ethyl acetate. The analysis of physico-chemical parameters indicated that all samples of plum seed oil have an exceptional quality. Schaal oven test indicated that the fixed oil of plum seed exhibited satisfactory oxidative stability at moderate storage temperatures (up to 65 °C). The composition of phenolic compounds in the oil samples was determined using HPLC method. The most abundant compound of seven identified and quantified phenolic compounds was vanillic acid. The highest content of ß-carotene (1.67 mg 100 g-1 fixed oil) spectrophotometrically determined was in the oil extracted with n-hexane. The lowest content of ß-carotene (1.26 mg 100 g-1 fixed oil) was determined in the oil extracted with a mixture of chloroform:methanol (2:1 v/v). This oil had the highest antioxidant activity (IC50 value of 4.35 mg mL-1) compared to other oil samples. The antioxidant activity was probably caused by the presence of phenolic compounds. The investigated physico-chemical properties demonstrated that the plum seed oil has a potential for application in the food, cosmetics, and pharmaceutical industry.

Drug design strategies with metal-hydroxyquinoline complexes.

Introduction: 8-hydroxyquinoline derivatives and their complexes with transition metals are the subject of many studies due to their anticancer, anti-inflammatory, anti-infective, and antidiabetic activities.Areas covered: Within this article, the authors review the synthesis and current applications of metal-8-hydroxyquinoline complexes in drug design with a critical overview of the latest advancements in the field.Expert opinion: Metal-8-hydroxyquinoline complexes are especially interesting because of their simple synthesis procedures and possible applications in modern medicine. The complexation between transition metal ions and 8-hydroxyquinoline or its derivatives is achieved via their O and N atoms. The main problem with their application is lipophilicity. This particular property has an impact on their solubility, biological activity, transport through the cell membrane, construction of the complex with a receptor, and development of drugs. Furthermore, in the treatment of neurodegenerative diseases and brain cancers, the passage of the complexes through the blood-brain barrier can only be ensured through novel drug design.

Optimization of Ultrasound-Assisted Extraction of Phenolic Compounds from Black Locust (Robiniae Pseudoacaciae) Flowers and Comparison with Conventional Methods.

The aim of this study was to optimize the ultrasound-assisted extraction of phenolic compounds from black locust (Robiniae pseudoacaciae) flowers using central composite design. The ethanol concentration (33-67%), extraction temperature (33-67 °C), and extraction time (17-33 min) were analyzed as the factors that impact the total phenolic content. The liquid-to-solid ratio of 10 cm3 g-1 was the same during extractions. The optimal conditions were found to be 59 °C, 60% (v/v) ethanol, and extraction time of 30 min. The total phenolic content (TPC = 3.12 gGAE 100 g-1 dry plant material) and antioxidant activity (IC50 = 120.5 µg cm-3) of the extract obtained by ultrasound-assisted extraction were compared with those obtained by maceration (TPC = 2.54 gGAE 100 g-1 dry plant material, IC50 = 150.6 µg cm-3) and Soxhlet extraction (TPC = 3.22 gGAE 100 g-1 dry plant material, IC50 = 204.2 µg cm-3). The ultrasound-assisted extraction gave higher total phenolic content and better antioxidant activity for shorter extraction time so that it represents the technique of choice for the extraction of phenolic compounds. The obtained extract, as the source of antioxidants, can be applied in the pharmaceutical and food industries.

The effect of complexation with cyclodextrins on the antioxidant and antimicrobial activity of ellagic acid.

PURPOSE: The aim of the paper was to develop the simple procedures for preparation of inclusion complexes of ellagic acid (EA) with cyclodextrins (CDs) and to investigate their antioxidant and antimicrobial activity. METHODS: The structural characterization was carried out using Fourier transform infrared spectroscopy (FTIR), X-ray diffraction (XRD), and nuclear magnetic resonance (NMR) methods. The phase solubility technique was used to investigate the interactions between 'host' and 'guest' molecules and to estimate the molar ratio between them. The antioxidant and antimicrobial activity of EA and inclusion complexes were determined. RESULTS: The apparent stability constants were found to be 117 dm3 mol-1 for the complex with ß-CD and 161 dm3 mol-1 for the complex with (2-hydroxypropyl)-ß-cyclodextrin (HP-ß-CD). The results of phase-solubility studies showed that EA formed the inclusion complexes with CDs in the molar ratio of 1:1. The calculated half-maximal inhibitory concentration was 41.18 µg cm-3 for butyl hydroxy toluene, 1.96 µg cm-3 for EA, 0.88 µg cm-3 for inclusion complex with HP-ß-CD, and 1.27 µg cm-3 for inclusion complex with ß-CD. CONCLUSION: The stability constants indicated the rapid release of EA from the inclusion complexes in the aqueous medium at 25 °C. The antioxidant activity of EA was increased, while the antimicrobial activity was preserved after complexation with CDs.

Preparation, characterization and antimicrobial activity of inclusion complex of biochanin A with (2-hydroxypropyl)-ß-cyclodextrin.

OBJECTIVES: An inclusion complex of biochanin A (BCA) with (2-hydroxypropyl)-ß-cyclodextrin (HP-ß-CD) was prepared in the ethanol solution to improve its water solubility. METHODS: Using the FTIR, 1 H-NMR, XRD, DSC and SEM methods, the structural characterization of the prepared complex was analysed. KEY FINDINGS: The phase-solubility study has shown that the solubility of BCA was increased twofold in 42% (v/v) ethanol solution after complexation with HP-ß-CD. The complex between BCA and HP-ß-CD was prepared in the molar ratio of 1 : 1. The antibacterial activity of the inclusion complex was investigated against the various bacteria, fungus and yeast using the microdilution method. The minimal inhibitory concentration values for the analysed strain of bacteria were in the range of 0.84-1.69 mg/cm3 , whereby the prepared inclusion complex exhibited less effect on the reduction of the number of Escherichia coli and Klebsiella pneumoniae species compared to pure BCA. The inclusion complex of BCA was significantly more active against Candida albicans than pure BCA. Biochanin A and its inclusion complex has not expressed the activity against Aspergillus niger. CONCLUSIONS: Based on the obtained results, it can be concluded that the antimicrobial activity of BCA was remained unchanged after complexation.

Characteristics and Distribution of Penile Lesions in Croatian Men with Laboratory-Confirmed Low-Risk and High-Risk Human Papillomavirus (HPV) Infection.

BACKGROUND: Studies that compare penoscopic evaluation with the results of molecular human papillomavirus (HPV) testing are scarce. This study assessed and compared both type and distribution of various penile lesions using penoscopy (peniscopy) technique in Croatian men with different, laboratory-confirmed HPV status. METHODS: From a large cohort of men attending an outpatient STD clinic for HPV testing, a total of 120 patients were randomly selected and grouped into those positive for low-risk HPV, high-risk HPV, both low-risk and highrisk HPV, and those with negative results. Samples for HPV-DNA detection were taken by penile brushing and tested with the hc2 HPV DNA Test using Hybrid Capture 2 technology. Lesions were observed by photocolposcope after the application of aqueous 5% acetic acid, and classified as flat (macular), papular, papillary, classical condyloma, PIN-suspicious and non-specific lesions. RESULTS: The results have shown that flat and non-specific acetowhite lesions were the most common overall. All groups differed significantly with respect to papular and papillary lesions. A combination of heterogeneous lesions (i.e., a mixed penoscopic pattern) was commonly observed in all HPV-positive groups, but was significantly lower in the HPV-negative group. A majority of lesions were located in the coronal sulcus, and the four groups differed with respect to the location only considering the lesions of the penile shaft. Distribution of lesions in different sites did not appear to be significantly different with respect to the group, although differences within the high-risk HPV group and the group with both low-risk and high-risk HPV were detected. CONCLUSIONS: Penoscopy is relevant, but not a conclusive diagnostic tool for differentiating HPV from non-HPV findings in men - thus combining it with an HPV DNA test represents a more reliable approach.

Prevalence of HPV Infection in Croatian Men during a 12-year Period: a Comparative Study of External Genital and Urethral Swabs.

AIM: The aim of the study was to compare the prevalence and distribution of both low-risk and high-risk HPV infection of the urethra and outer genital region in Croatian men. Thus far there is a consensus that sampling the coronal sulcus and glans of the penis is essential for adequately assessing HPV status in men but less agreement is noted for urethral sampling. METHODS: External genital brushing and urethral swabs were taken from 1,342 men during a 12-year period and tested with the hc2 HPV DNA Test using Hybrid Capture 2 technology. RESULTS: The overall prevalence of male HPV infection in this study was 36.66%. Infection with high-risk HPV types (44.72%) was significantly more frequent than infection with low-risk HPV types (28.86%) or co-infection with both low-risk and high-risk HPV types (26.42%). HPV was more frequently demonstrated in the outer genital area (58.33%) when compared to the sole infection of the urethra (17.89%) or infection of both genital sites (23.78%). CONCLUSIONS: Results from this study indicate high prevalence of HPV infection in men and suggest that optimal sampling method for the testing of men is the combination of external genital and urethral swabs. Further research about the proper collection of biological samples and testing methods for HPV detection in men is necessary since our future end-goal is to implement standardized guidelines on sampling and diagnostic testing of males.

Optimization of technological procedure for amygdalin isolation from plum seeds (Pruni domesticae semen).

The process of amygdalin extraction from plum seeds was optimized using central composite design (CCD) and multilayer perceptron (MLP). The effect of time, ethanol concentration, solid-to-liquid ratio, and temperature on the amygdalin content in the extracts was estimated using both mathematical models. The MLP 4-3-1 with exponential function in hidden layer and linear function in output layer was used for describing the extraction process. MLP model was more superior compared with CCD model due to better prediction ability. According to MLP model, the suggested optimal conditions are: time of 120 min, 100% (v/v) ethanol, solid-to liquid ratio of 1:25 (m/v) and temperature of 34.4°C. The predicted value of amygdalin content in the dried extract (25.42 g per 100 g) at these conditions was experimentally confirmed (25.30 g per 100 g of dried extract). Amygdalin (>90%) was isolated from the complex extraction mixture and structurally characterized by FT-IR, UV, and MS methods.

Design and optimization of drugs used to treat copper deficiency.

INTRODUCTION: Copper is an essential element in the human organism. Furthermore, copper deficiency is rare; however, the hematologic manifestations associated with copper deficiency, such as anemia, leukopenia, neutropenia, myeloneuropathy and osteoporosis, are well known. AREAS COVERED: The authors present an overview of the various commercially available drugs used in the treatment of copper deficiency. Furthermore, the authors offer a description of copper complexes, as potential pharmaceutically active compounds, that can be used in the design of new formulations with therapeutic potential. EXPERT OPINION: Progress in the synthesis of new metallo-organic complexes (such as the copper-pullulan complex) and the chelated form of copper have provided new avenues for drug design that combat copper deficiency. The copper-pullulan complex, as an active compound, has been designed in its solid dosage form, and its optimization in the treatment of copper deficiency has been furthered through advancements in experimental design methodology. The authors believe that the numerous ongoing studies, evaluating the synthesis of these complexes, should produce new additions to the copper deficiency therapeutic armamentarium in the future.

The use of color duplex ultrasound and magnetic resonance imaging in the dissolution of idiopathic recurrent priapism in patient with congenital penile curvature--a case report.

Priapism, penile erection characterized as prolonged and devoid of sexual stimulation or excitement is a rare condition. It is critical to distinguish between low- and high-flow priapism, because the treatment algorithm differs markedly for these 2 conditions. The diagnosis is made clinically and confirmed with color Doppler ultrasonographic imaging (CDUS). We present a 21 year old men with high-flow priapism and left lateral congenital penile curvature. A duplex Winter shunt procedure was employed with corporeal irrigation of heparin solution and adrenalin solution instillation, but the priapism returned 12 hours after. Following several days slow instillation of phenilephrine or adrenaline solution accompanied by oral flutamide therapy resulted in complete detumescence. We used both CDUS and magnetic resonance imaging (MRI) before and after treatment of priapism. Although CDUS has been the primary modality for cross-sectional imaging of the penis, the superior soft-tissue contrast and spatial resolution afforded by MRI provide an opportunity to advance imaging evaluation of this organ.

Application of mathematical modeling for the development and optimization formulation with bioactive copper complex.

New formulation for treatment a copper deficiency in human organism was developed and optimized by application of mathematical modeling. This formulation contained copper (II) complex with polysaccharide pullulan, as active substance. The binder concentration [polyvinyl pyrrolidone (PVP %)], the disintegrant concentration (corn starch %) and the resistance to crushing (hardness) were taken as independent variables. In vitro measured drug release characteristics of the tablets at pH 1.20 and 7.56 were studied as response variables. Initially, the created full factorial 2(3) model showed that the resistance to crushing has the most significant effect on copper (II) complex release from the formulation. Optimal tablet formulation F2, with lower Hardness (50 N), lower Starch (20.0%) and higher PVP (2.7%) concentrations, is selected using the partial least squares (PLS) regression modeling. The selected formulation F2 has expressed the best drug release profile at both pH (98.66% pH = 1.20; 93.35% pH = 7.56), and the lowest variation of tablets weight. The presented theoretical approach and created PLS model can be readily applied in future copper complexes studies and formulation design.

Renal cell carcinoma with concurrent urothelial carcinoma of urinary bladder and non-Hodgkin lymphoma.

We report a case of a 71-year-old male with multiple primary malignancies involving kidney and urinary bladder, combined with synchronous lymphoma. The patient was admitted to the hospital because of painless gross hematuria. Examination revealed tumor of the right kidney and papillary tumor in the urinary bladder and enlarged lymph nodes along aorta and inferior vena cava. Transurethral resection of bladder tumor (TUR), radical nephrectomy of the right kidney and retroperitoneal lymphadenectomy were performed. Pathohistologic evaluation, together with immunohistochemistry, gave the patient the final diagnosis of renal cell carcinoma (RCC), urothelial carcinoma of the urinary bladder and B- small cell Non-Hodgkin lymphoma (B-CLL).

Development and optimization of formulation for treatment of copper deficiency in human organism.

The aim of this study was to design and optimize a new tablet formulation for treatment of copper deficiency in human organism by using an experimental design. The new no-veneered tablets, prepared by a wet granulation technique, are containg active substance, a copper(II) complex with polysaccharide pullulan. The binder concentration, the disintegrant concentration and the resistance to crushing were used as independent variables in the formulation, while in vitro measured drug release characteristics of the tablets was response variable in a full factorial design 2(3) modeling. A cubic model for data fitted was used to examine the obtained results. They showed that the resistance to crushing has the most significant effect on copper(II) complex release from the formulation, while the disintegrant concentration has smaller influence on dissolution profile of copper(II) complex and the binder concentration had minor impact in this study. Lower value of resistance to crushing has influence on better dissolution profile. Furthermore, physical characteristics of the tablets were evaluated, viz., drug content, hardness, thickness, friability, disintegration time, mass variation, particle size and size distribution.