Opioid Growth Factor and its Derivatives as Potential Non-toxic Multifunctional Anticancer and Analgesic Compounds.

Despite significant research progress on the pathogenesis, molecular biology, diagnosis, treatment, and prevention of cancer, its morbidity and mortality are still high around the world. The emerging resistance of cancer cells to anticancer drugs remains still a significant problem in oncology today. Furthermore, an important challenge is the inability of anticancer drugs to selectively target tumor cells thus sparing healthy cells. One of the new potential options for efficient and safe therapy can be provided by opioid growth factor (OGF), chemically termed Met-enkephalin. It is an endogenous pentapeptide (Tyr-Gly-Gly-Phe-Met) with antitumor, analgesic, and immune-boosting properties. Clinical trials have demonstrated that OGF therapy alone, as well as in combination with standard chemotherapies, is a safe, non-toxic anticancer agent that reduces tumor size. In this paper, we review the structure-activity relationship of OGF and its analogues. We highlight also OGF derivatives with analgesic, immunomodulatory activity and the ability to penetrate the blood-brain barrier and may be used as safe agents enhancing chemotherapy efficacy and improving quality of life in cancer patients. The reviewed papers indicate that Met-enkephalin and its analogues are interesting candidates for the development of novel, non-toxic, and endowed with an analgesic activity anticancer drugs. More preclinical and clinical studies are needed to explore these opportunities.

Effect of different conditions of synthesis on properties of silver nanoparticles stabilized by nanocellulose from carrot pomace.

The silver nanoparticles (AgNPs) can exhibit different optical properties depending on their size and shape as a result of synthesis method and the stabilizer used. In this research the synthesis of AgNPs in the presence of nanocellulose obtained from carrot pomace was investigated. The influence of silver nitrate concentration, temperature and mechanical agitation on size and shape of AgNPs was studied. The mixing of reagents during synthesis, regardless temperature, led to obtain AgNPs of various sizes and shapes. It was confirmed by different colors of samples with absorbance maximum from 334 to 779 nm, the transmission electron microscopy images and dynamic light scattering results. In unmixed samples only spherical nanoparticles with absorbance maximum at 408 nm were observed. Obtained results have demonstrated that mechanical agitation and an appropriate silver nitrate concentration combined with stabilizing effect of nanocellulose allow to obtain AgNPs in different shapes and sizes.

Evaluation of Nanocomposite Made of Polylactic Acid and Nanocellulose from Carrot Pomace Modified with Silver Nanoparticles.

In this research, it was proposed to use carrot cellulose nanofibrils (CCNF) isolated from carrot pomace modified with silver nanoparticles (AgNPs) as a filler of polylactic acid (PLA) composites matrix. The new procedure was based on two steps: first, the preparation of nanocellulose modified with metal nanoparticles, and then the combination with PLA. Two concentrations-0.25 mM and 2 mM-of AgNO3 were used to modify CCNF. Then, PLA was mixed with the filler (CCNF/AgNPs) in two proportions 99:1 and 96:4. The influence of CCNF/AgNPs on mechanical, hydrophilic, thermal, and antibacterial properties of obtained nanocomposites was evaluated. The greatest improvement of mechanical properties was observed for composite containing CCNF with 2 mM of AgNPs, which obtained the lowest Young modulus and highest strain at break. The degradation temperature was lower for PLA with CCNF/AgNPs, but crystallization temperature wasn't influenced. The addition of CCNF/AgNPs also increased hydrophilicity. The transmission rates of oxygen, nitrogen, and carbon dioxide also increased after the addition of CCNF/AgNPs to PLA. The antibacterial function against Escherichia coli and Bacillus cereus was obtained after the addition of AgNPs but only at the contact surface with the material made, suggesting the lack of migration of nanoparticles from the composite.

Enzymes and vitamin C as factors influencing the presence of arabinogalactan proteins (AGPs) in Solanum lycopersicum fruit.

Arabinogalactan proteins (AGPs) are ubiquitous components of the amorphous plant extracellular matrix. They are characterized by a high proportion of sugar moieties, heterogeneity of their protein backbone and carbohydrate chains. It is known that AGPs form a complex network with other basic constituents in cell wall thus it may also play a role in softening process of fruit. The use of enzymatic degradation and cell wall polysaccharide directed probes are valid analytical tools for the study of developmental modification of the fruit structure. However, it is unknown whether pectolytic enzymes affect AGPs. Thus, the aim of the current work is to detect AGP epitopes in situ to understand the impact of selected degradation enzymes on various carbohydrate moieties of AGPs. Secondly, there are no data with clarification of the impact of vitamin C on fruit ripening processes at the cellular level; hence, we also focused on the effect of vitamin C on the arrangement of AGPs as important constituents of the polysaccharide-proteoglycan network in the fruit cell wall. The results indicate that the distribution of the examined AGP carbohydrate moieties differs, which are related to changes in tissue architecture. The absence of glycan chains causes disruption in establishment of correlations between cell wall constituents and rearrangement in the cell wall structure. The induced modifications of cell walls are not comparable to alterations occurring in naturally ripening fruit, which allows a conclusion that the synergistic action of a wide variety of factors influences ripening.

Tailored nanocellulose structure depending on the origin. Example of apple parenchyma and carrot root celluloses.

Cellulose is the major polysaccharide of cell walls in every plant, making it one of the most abundant natural polymers on Earth. However, despite many decades of investigations, the supramolecular structure of cellulose and especially its variation in the cell walls of different plants have still not been fully revealed. In the present study, cellulose from the parenchymatic tissue of apple fruits and carrot roots was isolated, and nanocellulose was further prepared by high-intensity ultrasonication. AFM revealed that the obtained nanocellulose differed in dimension between the two plant species. Compared with carrot cellulose, whose nanocellulose was obtained in the form of whiskers, apple cellulose had longer and thinner nanofibrils. Both nanocellulose types also differed in terms of their crystalline structure. XRD data indicated that, compared with the apple cellulose, the carrot cellulose had a higher degree of crystallinity and larger crystallites. Moreover, FTIR and Raman spectroscopy revealed differences between the cellulose types in terms of their methine environment, hydroxymethyl conformations and skeletal vibrations. Additionally, with respect to their mechanical properties, the less crystalline apple cellulose and nanocellulose films were more elastic than the stiffer carrot cellulose and nanocellulose films. The possible reason for such differences between the two cellulose types is related to differences in plant tissue morphology and function. During development, apple fruit cell walls must withstand increasing turgor, probably higher that in the case of carrot tissue; therefore, the cellulose scaffolding must be elastic and strong. On the other hand, carrot, a root vegetable, also has to be strong enough to penetrate the soil as well as for its own growth; thus, the cell wall and cellulose scaffold have to be stiff and tough. Thus the structure of nanocellulose depends not only on the treatment but also on the cellulose source.

Distribution of arabinogalactan proteins and pectins in the cells of apple (Malus × domestica) fruit during post-harvest storage.

Background and Aims: Changes in the arrangement of cell wall components determine cell wall properties (integrity, stiffness), thereby affecting the macro-scale properties of fruits, which are important for consumers and industry. Arabinogalactan proteins (AGPs) are ubiquitous components of the plant cell, in which they have various functions. Currently, AGPs are considered to be one of the less well-known, enigmatic proteoglycans, a consequence of their heterogeneous structure and unclear mechanism of activity. Methods: An immunocytochemical study was conducted to elucidate the distribution of AGPs and pectic polysaccharides contained in apple (Malus × domestica) fruit during senescence. De-esterified homogalacturonan (LM19), methyl-esterified homogalacturonan (LM20), processed arabinan (LM16) and three AGP epitopes (JIM13, JIM15, MAC207) were identified in the fruit at three stages: fresh fruit, and fruit at 1 and 3 months of post-harvest storage. Key Results: Microscopy revealed spatio-temporal changes in the localization of all examined epitopes. Changes of fruit cell wall assembly and its degradation were confirmed by determination of the galacturonic acid content in the WSP (water soluble pectins), CSP (chelator soluble pectins) and DASP (dilute alkali soluble pectins) fractions. Conclusions: The results revealed dependencies between AGPs, arabinan and homogalacturonan distribution in apple fruit, which are correlated with changes in microstructure during senescence. We propose that AGPs are involved in establishment of the cell wall - plasma membrane continuum.

Structural network of arabinogalactan proteins (AGPs) and pectins in apple fruit during ripening and senescence processes.

The cell wall is an essential framework determining the overall form of the plant cell. Our study was focused on the distribution of arabinogalactan proteins (AGPs), arabinan, and homogalacturonan in fruit cells during ripening and storage with emphasis on quantitative analysis of their presence in particular regions of the cell wall - plasma membrane. The localization of the examined compounds was determined with immunohistochemistry techniques and immunogold labelling. Spatio-temporal colocalization between AGPs epitopes - [ßGlcA(1→3)-αGalA(1→2)Rha] recognized by JIM13 and MAC207 antibodies, and arabinan labelled by the LM16 antibody was detected in the inner cell wall layer, in association with the plasma membrane. The specific arrangement of AGP and arabinan epitopes differentiated them from homogalacturonan epitopes, consisting of GalA residues recognized by LM19 and LM20 antibodies in all the examined fruit maturity stages. The disruption of cell wall - plasma membrane continuum, observed during ripening-associated softening process, was associated with both the substantial decrease of AGPs, pectins content and with remodeling of their arrangement. The results indicate that the textural properties of fruit during growth and postharvest storage, an attribute of fruit quality becoming selection criteria for consumers, depend on the existence of dynamic network organizing polysaccharides and glycoproteins in the extracellular matrix.

Changing of biochemical parameters and cell wall polysaccharides distribution during physiological development of tomato fruit.

This study was aimed at discovering an impact of biochemical parameters (like content of cell wall polysaccharides, phenolic compounds, ascorbic acid or activity of pectinolytic enzymes) on cell wall microstructure during physiological fruit development. Cell wall microstructure as well as changes in the polysaccharides distribution were examined by confocal Raman microscopy. Also there was a need to simultaneous usage of reference method which is immunolabeling. A tomato fruit (Solanum lycopersicum cv Cerise) has been selected to observe the changes taking place in the fruit cell wall as it recently has been recognized as a model species for exploring fruit development processes such as fruit formation and ripening. Our studies showed that chemical images allows to depict changes in spatial distribution of polysaccharides in plant cell wall (including the middle lamella area), thus this technique allows to observation of cell wall degradation during tomato ripening (mainly pectic polysaccharides degradation). It seems that high level of pectinolytic enzymes activity and increasing content of ascorbate and hence decrease of pectins content have a significant impact on spatial distribution of biopolymers in fruit cell wall.

Simultaneous influence of pectin and xyloglucan on structure and mechanical properties of bacterial cellulose composites.

The impact of the matrix polysaccharides on the cellulose microfibrils structure as well as on the mechanical properties of cell walls still remains an open question. Therefore, the aim of investigations was to determine the simultaneous influence of (i) different concentrations of pectins with constant concentration of xyloglucan, and (ii) different concentrations of xyloglucan with constant concentration of pectins on cellulose structure. Composites of bacterial cellulose (BC) produced by Komagataeibacter xylinus are considered to mimic natural plant cell walls. This investigation showed that the lower the ratio of xyloglucan to pectin was, the higher Young's modulus of BC composite was and also obtained cellulose microfibrils were thinner. The increasing concentration of xyloglucan to pectin also caused the drop down in microfibrils crystallinity degree with predominant structure of cellulose Iß. In that case, also the length of cellulose chains was growing and reaching the highest value among all BC composites.

Isolation and Characterization of Cellulose from Different Fruit and Vegetable Pomaces.

A new fractionation process was developed to achieve valorization of fruit and vegetable pomaces. The importance of the residues from fruits and vegetables is still growing; therefore; the study presents the novel route of a fractioning process for the conversion of agro-industrial biomasses, such as pomaces, into useful feedstocks with potential application in the fields of fuels, chemicals, and polymers. Hence, the biorefinery process is expected to convert them into various by-products offering a great diversity of low-cost materials. The final product of the process is the cellulose of the biofuel importance. The study presents the novel route of the fractioning process for the conversion of agro-industrial biomasses, such as pomaces, into useful feedstocks with a potential application in the fields of fuels, chemicals, and polymers. Therefore the aim of this paper was to present the novel route of the pomaces fraction and the characterization of residuals. Pomaces from apple, cucumber, carrot, and tomato were treated sequentially with water, acidic solution, alkali solution, and oxidative reagent in order to obtain fractions reach in sugars, pectic polysaccharides, hemicellulose, cellulose, and lignin. Pomaces were characterized by dry matter content, neutral detergent solubles, hemicellulose, cellulose, and lignin. Obtained fractions were characterized by the content of pectins expressed as galacturonic acid equivalent and hemicelluloses expressed as a xyloglucan equivalent. The last fraction and residue was cellulose characterized by crystallinity degree by X-ray diffractometer (XRD), microfibril diameter by atomic force microscope (AFM), and overall morphology by scanning electron microscope (SEM). The hemicelluloses content was similar in all pomaces. Moreover, all the materials were characterized by the high pectins level in extracts evaluated as galacturonic acid content. The lignins content compared with other plant biomasses was on a very low level. The cellulose fraction was the highest in cucumber pomace. The cellulose fraction was characterized by crystallinity degree, microfibril diameter, and overall morphology. Isolated cellulose had a very fine structure with relatively high crystalline index but small crystallites.

FT-IR and FT-Raman characterization of non-cellulosic polysaccharides fractions isolated from plant cell wall.

The purpose of this work was to reveal the structural changes of cell wall polysaccharides' fractions during tomato fruit development by analysis of spectral data. Mature green and red ripe tomato fruit were taken into consideration. The FT-IR spectra of water soluble pectin (WSP), imidazole soluble pectin (ISP) and diluted alkali soluble pectin (DASP) contained bands typical for pectins. Whereas for KOH fraction spectra bands typical for hemicelluloses were present. The FT-IR spectra showed the drop down of esterification degree of WSP and ISP polysaccharides during maturation. The changes in polysaccharides structure revealed by spectra were the most visible in the case of pectic polysaccharides. The WSP and DASP fraction pectins molecules length were shortened during tomato maturation and ripening. Whereas the ISP fraction spectra analysis showed that this fraction contained rhamnogalacturonan I, but also for red ripe was rich in pectic galactan comparing with ISP fraction from mature green.

Determination of the Optimum Harvest Window for Apples Using the Non-Destructive Biospeckle Method.

Determination of the optimum harvest window plays a key role in the agro-food chain as the quality of fruit depends on the right harvesting time and appropriate storage conditions during the postharvest period. Usually, indices based on destructive measurements are used for this purpose, like the De Jager Index (PFW-1), FARS index and the most popular Streif Index. In this study, we proposed a biospeckle method for the evaluation of the optimum harvest window (OHW) of the "Ligol" and "Szampion" apple cultivars. The experiment involved eight different maturity stages, of which four were followed by long cold storage and shelf life to assist the determination of the optimum harvest window. The biospeckle activity was studied in relation to standard quality attributes (firmness, acidity, starch, soluble solids content, Streif Index) and physiological parameters (respiration and ethylene emission) of both apple cultivars. Changes of biospeckle activity (BA) over time showed moderate relationships with biochemical changes during apple maturation and ripening. The harvest date suggested by the Streif Index and postharvest quality indicators matched with characteristic decrease in BA. The ability of biospeckle method to characterize the biological state of apples was confirmed by significant correlations of BA with firmness, starch index, total soluble solids and Streif Index, as well as good match with changes in carbon dioxide and ethylene emission. However, it should be noted that correlations between variables changing over time are not as meaningful as independent observations. Also, it is a well-known property of the Pearson's correlation that its value is highly susceptible to outlier data. Due to its non-selective nature the BA reflected only the current biological state of the fruit and could be affected by many other factors. The investigations showed that the optimum harvest window for apples was indicated by the characteristic drop of BA during pre-harvest development. Despite this, at the current state of development the BA method cannot be used as an indicator alone. Due to rather poor results for prediction in OHW the BA measurements should be supported by other destructive methods to compensate its low selectivity.

Raman imaging of changes in the polysaccharides distribution in the cell wall during apple fruit development and senescence.

MAIN CONCLUSION: Du ring on-tree ripening, the pectin distribution changed from polydispersed in cell wall to cumulated in cell wall corners. During apple storage, the pectin distribution returned to evenly dispersed along the cell wall. The plant cell wall influences the texture properties of fruit tissue for example apples become softer during ripening and postharvest storage. This softening process is believed to be mainly connected with changes in the cell wall composition due to polysaccharides undergoing an enzymatic degradation. These changes in polysaccharides are currently mainly investigated via chemical analysis or monoclonal labeling. Here, we propose the application of Raman microscopy for evaluating the changes in the polysaccharide distribution in the cell wall of apples during both ripening and postharvest storage. The apples were harvested 1 month and 2 weeks before optimal harvest date as well as at the optimal harvest date. The apples harvested at optimal harvest date were stored for 3 months. The Raman maps, as well as the chemical analysis were obtained for each harvest date and after 1, 2 and 3 months of storage, respectively. The analysis of the Raman maps showed that the pectins in the middle lamella and primary cell wall undergo a degradation. The changes in cellulose and hemicellulose were less pronounced. These findings were confirmed by the chemical analysis results. During development changes of pectins from a polydispersed form in the cell walls to a cumulated form in cell wall corners could be observed. In contrast after 3 months of apple storage we could observe an substantial pectin decrease. The obtained results demonstrate that Raman chemical imaging might be a very useful tool for a first identification of compositional changes in plant tissue during their development. The great advantage Raman microspectroscopy offers is the simultaneous localization and identification of polysaccharides within the cell wall and plant tissue.

Study on dietary fibre by Fourier transform-infrared spectroscopy and chemometric methods.

Fresh fruit is an important part of the diet of people all over the world as a significant source of water, vitamins and natural sugars. Nowadays it is also one of the main sources of dietary fibre. In fruit the dietary fibre is simply cell wall consisting essentially of polysaccharides. The aim of present study was to predict the contents of pectins, cellulose and hemicelluloses by partial least squares regression (PLS) analysis on the basis of Fourier transform-infrared (FT-IR) spectra of fruit cell wall residue. The second purpose was to analyse the composition of dietary fibre from fruit based on FT-IR spectral information in combination with chemometric methods (principle components analysis (PCA) and hierarchical cluster analysis (HCA)). Additionally the contents of polysaccharides in studied fruits were determined by analytical methods. It has been shown that the analysis of infrared spectra and the use of multivariate statistical methods can be useful for studying the composition of dietary fibre.

Imaging of polysaccharides in the tomato cell wall with Raman microspectroscopy.

BACKGROUND: The primary cell wall of fruits and vegetables is a structure mainly composed of polysaccharides (pectins, hemicelluloses, cellulose). Polysaccharides are assembled into a network and linked together. It is thought that the percentage of components and of plant cell wall has an important influence on mechanical properties of fruits and vegetables. RESULTS: In this study the Raman microspectroscopy technique was introduced to the visualization of the distribution of polysaccharides in cell wall of fruit. The methodology of the sample preparation, the measurement using Raman microscope and multivariate image analysis are discussed. Single band imaging (for preliminary analysis) and multivariate image analysis methods (principal component analysis and multivariate curve resolution) were used for the identification and localization of the components in the primary cell wall. CONCLUSIONS: Raman microspectroscopy supported by multivariate image analysis methods is useful in distinguishing cellulose and pectins in the cell wall in tomatoes. It presents how the localization of biopolymers was possible with minimally prepared samples.