Chemopreventive properties of spent hops (Humulus Lupulus L.) extract against angiogenesis, invasion and migration of colorectal cancer cells.

There is a great deal of interest in identifying new chemopreventive agents for colorectal cancer, one of the leading causes of cancer-related death. One promising group of candidates is the polyphenols; being natural compounds with high structural diversity, they have a very wide spectrum of anti-inflammatory, anti-oxidant and anti-cancer properties. The present study reports for the first time that spent hops extract (SHE) inhibits the angiogenesis, invasion and migration of SW-480 and HT-29 colorectal adenocarcinoma cells; after incubation with 200 µg/mL SHE, SW-480 and HT-29 cell invasion fell by 98.5% and 89% vs. controls, and migration was inhibited by 99% and 88% vs. controls. These changes were accompanied by a decline of matrix metalloproteinase 2 (MMP-2) and MMP-9 expression and activity. In addition, SHE reduced the expression of vascular endothelial growth factor and hypoxia-inducible factor 1α for both cell lines, indicating that the tested extract has anti-angiogenic potential. In conclusion, our data shows that SHE may be an effective chemopreventive agent acting via the inhibition of angiogenesis, invasion and migration of colorectal cancer cells.

Valorisation of agri-food waste to fertilisers is a challenge in implementing the circular economy concept in practice.

The area of agricultural wastes valorisation to fertilizers is attracting growing attention because of the increasing fertilizer prices of fertilizers and the higher costs of waste utilization. Despite the scientific and political interest in the concept of circular economy, few studies have considered the practical approach towards the implementation of elaborated technologies. This article outlines innovative strategies for the valorisation of different biobased wastes into fertilizers. The present work makes a significant contribution to the field of new ideas for waste biomass management to recover significant fertilizer nutrients. These results emphasize the importance of the biomass use as a base of renewable resources, which has recently gained special importance, especially in relation to the outbreak of pandemia and war. Broken supply chains and limited access to deposits of raw materials used in fertilizer production (natural gas, potassium salts) meant that now, as never before, it has become more important and feasible to implement the idea of a circular economy and a green deal. We have obtained satisfactory results that demonstrate that appropriate management of biological waste (originating from agriculture, food processing, aquaculture, forest, pharmaceutical industry, and other branches of industry, sewage sludge) will not only reduce environmental nuisance (reducing waste heaps), but will also allow recovery of valuable materials, such as nitrogen (especially valuable amino acids), phosphorus, potassium, microelements, and biologically active substances with properties that stimulate plant growth. The results reported here provide information on production of biobased plant protection products (bioagrochemicals) from agri-food waste. This work reports an overview of biopesticides and biofertilisers production technologies and summarizes their properties and the mechanisms of action.

Chemoprotective effects of Japanese quince (Chaenomeles japonica L.) phenol leaf extract on colon cancer cells through the modulation of extracellular signal-regulated kinases/AKT signaling pathway.

Japanese quince leaf phenol-rich extract (PRE) represents a good source of phenolic compounds, among which chlorogenic acid and naringenin hexoside are the main constituents. The aim of this research was to evaluate the chemopreventive activity of PRE in human colon cancer (SW-480 and HT-29) and human normal colon cell line (CCD 841 CoN). All cell lines were exposed to different concentrations of the extract (150-500 κg/mL for SW-480 and CCD 841 CoN; and 250-750 κg/mL for HT-29) to investigate migration and invasion, as well as the activity and secretion of metalloproteinases (MMP-2 and MMP-9) involved in these mechanisms. Moreover, the influence of PRE on the activity of ERK and AKT pathways, which are strongly involved in colon cancer development (CRC), were measured. Our results demonstrated that PRE significantly inhibited migration and invasion in SW-480, HT-29 and CCD 841 CoN cells through MMP-2 and MMP-9-dependent mechanisms. We also proved that PRE can effectively downregulate both the activity and protein expression of MMP-2 and MMP-9 in these cell lines. The exception was the higher concentration of PRE, which up-regulated the protein expression of MMP-9 in SW-480. Additionally, we showed that significant inhibition of p-ERK/p-AKT expression in SW-480 after treatment with PRE is involved in chemopreventive effects of this extract. In case of exposure of HT-29 cells to PRE, we observed a significant upregulation of p-ERK protein expression, and suppression of p-AKT mechanism. This research of Japanese quince phenol leaf extract suggests its application in colon cancer prevention and treatment due to its ability to inhibit migration and invasion in MMP-9 and MMP-2-dependent mechanisms via most likely the modulation of ERK and AKT signaling pathways in colon cancer cells. Overall, our results provide an experimental foundation for further research on its potential activities and effects in vivo.

An examination of maximum legal application rates of dairy processing and associated STRUBIAS fertilising products in agriculture.

The dairy industry produces vast quantities of dairy processing sludge (DPS), which can be processed further to develop second generation products such as struvite, biochars and ashes (collectively known as STRUBIAS). These bio-based fertilizers have heterogeneous nutrient and metal contents, resulting in a range of possible application rates. To avoid nutrient losses to water or bioaccumulation of metals in soil or crops, it is important that rates applied to land are safe and adhere to the maximum legal application rates similar to inorganic fertilizers. This study collected and analysed nutrient and metal content of all major DPS (n = 84) and DPS-derived STRUBIAS products (n = 10), and created an application calculator in MS Excel™ to provide guidance on maximum legal application rates for ryegrass and spring wheat across plant available phosphorus (P) deficient soil to P-excess soil. The sample analysis showed that raw DPS and DPS-derived STRUBIAS have high P contents ranging from 10.1 to 122 g kg-1. Nitrogen (N) in DPS was high, whereas N concentrations decreased in thermo-chemical STRUBIAS products (chars and ash) due to the high temperatures used in their formation. The heavy metal content of DPS and DPS-derived STRUBIAS was significantly lower than the EU imposed limits. Using the calculator, application rates of DPS and DPS-derived STRUBIAS materials (dry weight) ranged from 0 to 4.0 tonnes ha-1 y-1 for ryegrass and 0-4.5 tonnes ha-1 y-1 for spring wheat. The estimated heavy metal ingestion to soil annually by the application of the DPS and DPS-derived STRUBIAS products was lower than the EU guideline on soil metal accumulation. The calculator is adaptable for any bio-based fertilizer, soil and crop type, and future work should continue to characterise and incorporate new DPS and DPS-derived STRUBIAS products into the database presented in this paper. In addition, safe application rates pertaining to other regulated pollutants or emerging contaminants that may be identified in these products should be included. The fertilizer replacement value of these products, taken from long-term field studies, should be factored into application rates.

The influence of polyphenol-rich extracts on the production of pro-inflammatory mediators in macrophages.

Recent decades have seen a rise in chronic inflammatory diseases such as diabetes, cardiovascular diseases, asthma, rheumatoid arthritis, neurodegenerative diseases. Importantly, such chronic inflammatory diseases also increase the risk of cancer development and there is a pressing need to identify new anti-inflammatory drugs. One promising source of new medication are natural polyphenolic compounds and polyphenol-rich preparations, extracts and foods, which have strong antioxidant properties. This paper reviews the anti-inflammatory role of polyphenolic-rich natural extracts, and their ability to modulate crucial pro-inflammatory mediators, such as cyclooxygenase-2, prostaglandin E2, inducible nitric oxide synthase, and nitric oxide, in macrophage cells. Our research confirms that natural compounds have health potential, and could be used in the treatment or prevention of inflammatory diseases.

Phytochemicals containing biologically active polyphenols as an effective agent against Covid-19-inducing coronavirus.

The outbreak of Covid-19 disease caused by SARS-CoV-19, along with the lack of targeted medicaments and vaccines, forced the scientific world to search for new antiviral formulations. In this review, we describe the current knowledge about plant extracts containing polyphenols that inhibit Covid-19. Many plant-derived natural compounds (polyphenols) might provide a starting point for the research on the use of plant extracts in coronavirus treatment and prevention. Antivirus polyphenolic drugs can inhibit coronavirus enzymes, which are essential for virus replication and infection. This group of natural substances (betulinic acid, indigo, aloeemodine, luteolin, and quinomethyl triterpenoids, quercitin or gallates) is a potential key to designing antiviral therapies for inhibiting viral proteases. The known pharmacophore structures of bioactive substances can be useful in the elaboration of new anti-Covid-19 formulations. The benefit of using preparations containing phytochemicals is their high safety for patients and no side effects.

Comparison of phenolic compounds, antioxidant and cytotoxic activity of extracts prepared from Japanese quince (Chaenomeles japonica L.) leaves.

Phenolic compounds are very important in the prevention and treatment of many civilization diseases, including colorectal cancer (CRC). In the present study we investigated and compared the phytochemical composition, antioxidant and cytotoxic activity of Japanese quince (Chaenomeles japonica L.) leaves crude phenolic extract (CPE) and purified phenolic-rich extracts (PRE). The UPLC-Q-TOF-MS analysis showed that both extracts contain diversified phenolics compounds (33 - 36 compounds in the PRE and CPE, respectively), among which chlorogenic acid and naringenin hexoside turned out to be the main constituents. Both FRAP and ABTS tests showed that PRE had 2-fold higher antioxidant activity compared to CPE. Furthermore, PRE exhibited a higher cytotoxic activity towards colon cancer cells (SW-480 and HT-29) than CPE. After 24-hours incubation with PRE the IC50 value for SW-480 cell line was obtained at the concentration of 239 µg/mL, while CPE treatment caused the same decrease only after 72h at 277 µg/mL. In addition, PRE had a stronger cytotoxic effect on the colon cancer cell lines (SW-480 and HT-29) than on normal intestinal cells (CCD-18Co and CCD 841 CoN). These results provide the first evidence that extracts from Japanese quince leaves (especially phenolic-rich extract, PRE) strongly decrease the viability of both SW-480 and HT-29 lines, which may suggest their cytotoxic activity towards colon cancer cells.

Spent hops (Humulus Lupulus L.) extract as modulator of the inflammatory response in lipopolysaccharide stimulated RAW 264.7 macrophages.

Macrophages play important roles in acute and chronic inflammation. Upon their activation, they secrete a variety of mediators, including eicosanoids, nitric oxide and cytokines, which play different roles in the stimulation and resolution of inflammatory processes. There is a continuous search for selective modulators of these processes. Natural polyphenols and polyphenol-rich extracts have been found to possess preventive and therapeutic potential, including by their anti-inflammatory effects. In this study, the inhibition of the formation of inflammatory mediators by the spent hops extract (SHE), a polyphenol-rich extract from Humulus Lupulus L., was examined using lipopolysaccharide (LPS)- activated murine macrophages (RAW 264.7). The SHE suppressed inter alia the interleukin-6 (IL-6) mRNA expression to 32% in LPS-activated macrophages and to 61% at a protein level (at 25 µg/mL). SHE reduced both the cyclooxygenase-2 (COX-2) mRNA expression to 47% and their protein expression to 32%. Not only did SHE inhibit the IL-6 and COX-2 levels but also decreased both inducible nitric oxide synthase (iNOS) protein expression to 2% at 25 µg/mL and nitric oxide (NO) production for all tested concentrations. The inhibited expression of these inflammatory molecules was likely caused by the reduced activity of nuclear factor-κB (NF-κB). Both mRNA and protein expression of NF-κB was decreased to 38% and 42%, respectively. These results provide the first evidence that SHE decreases the expression of proinflammatory cytokines and inflammatory mediators, which merits further studies to investigate the potential of SHE as anti-inflammatory preparation.

Japanese quince (Chaenomeles japonica) leaf phenol extract as modulator of the inflammatory response in lipopolysaccharide-triggered murine macrophage RAW 264.7 cells.

In the present study we investigated the anti-inflammatory activity of Japanese quince leaf polyphenol-rich extract (JQLPE) in lipopolysaccharide (LPS)-activated murine macrophages (RAW 264.7). The Q-PCR analysis revealed that JQLPE decreased Nfkb1, Ptgs2, and Il1b expression at the mRNA level by 80%, 50% and 48%, respectively. Similarly, JQLPE significantly attenuated expression of inducible nitric oxide synthase (iNOS), cyclooxygenase 2 (COX-2), interleukin-1beta (IL-1ß), IL-6 and tumor necrosis factor alpha (TNF-α) (by 60%, 50%, 67%, 37% and 36%, respectively) at the protein level and nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW264.7. Western blot also showed that the expression of nuclear factor kappaB (NF-κB) p65 and p-NF-κB p65 was down-regulated after JQLPE treatment. These results provide the first evidence that JQLPE decreases the expression of pro-inflammatory cytokines (IL-1ß, IL-6, TNF-α), inflammatory mediators (COX-2, iNOS) and both NF-κB p65 and p-NF-κB p65 in LPS-stimulated RAW264.7 cells, which may suggest its anti-inflammatory activity.

Potential applications of cyanobacteria: Spirulina platensis filtrates and homogenates in agriculture.

In the present paper, products obtained from a blue-green microalga Spirulina platensis filtrate (applied for seed soaking and for foliar spray) and homogenate (used for seed coating) were tested in the cultivation of radish. Their effect on length, wet mass, multielemental composition and the greenness index of the radish leaves was examined. Multi-elemental analyses of the algal products, and radish were also performed using inductively coupled plasma-optical emission spectrometry (ICP-OES). The best soaking time, concentrations of filtrate and doses of homogenate were established. The longest and heaviest plants were observed for homogenate applied at a dose of 300 µL per 1.5 g of seeds and 15% of filtrate applied as foliar spray. The highest chlorophyll content was found in the group treated with 100 µL of homogenate and 5% of filtrate. In the case of soaking time, the longest plants were in the group where seeds were soaked for 6 h, but the heaviest and greenest were after soaking for 48 h. The applied algal products increased the content of elements in seedlings. Obtained results proved that algal extracts have high potential to be applied in modern horticulture and agriculture. The use of Spirulina-based products is consistent with the idea of sustainable agriculture that could help to ensure production of sufficient human food to meet the needs of rising population and protection of the environment.

Soybean meal enriched with microelements by biosorption--a new biological feed supplement for laying hens. Part I. Performance and egg traits.

The aim of the study was to evaluate the effect of soybean meal enriched with Cu(II), Zn(II), Fe(II) and Cr(III) by biosorption on egg traits (egg weight, eggshell strength, eggshell thickness, yolk colour, albumen height) and performance of laying hens. Also, the effect of increased microelement doses in biological form on egg quality parameters and hens performance was investigated. A consumer questionnaire was undertaken to evaluate the organoleptic parameters of the eggs. Generally, our study showed that in the groups fed with the new biological supplement, egg quality parameters improved, including eggshell strength, eggshell thickness, albumen height and yolk colour. The biological form of microelements also improved the feed conversion rate, especially in the group fed with a biological form of Cr(III). Moreover, the new supplement improved organoleptic parameters of the eggs, in comparison to the inorganic form of microelements as well as to chelate. Enriched soybean meal could constitute an alternative for currently used feed additives with microelements.

Effect on supplementation of Spirulina maxima enriched with Cu on production performance, metabolical and physiological parameters in fattening pigs.

In this paper, the effect of addition of the biomass of Spirulina maxima enriched with copper (Sm-Cu) to the animal feed is discussed. The biomass was cultivated in the photobioreactor with the capacity of 10 m3. After the biosorption process, the enriched biomass was investigated as the source of valuable nutrients. The feeding experiment was conducted for 87 days. The study was performed in individual rearing pens, with controlled microclimate, feed and water were available semi-ad libitum. Piglets (24) were divided into two groups (control and experimental). The experimental group was fed with addition of the biomass of Sm-Cu instead of inorganic salts. There were no statistically significant differences between the average daily and periodic weight gain, daily and periodic feed collection, as well as feed conversion ratio. There were no statistically significant differences between the amount of N excreted in faeces and urine, when considering the retention of N, both in relation to the consumed N, and relative N digested which was at a similar level. In the experimental group in comparison to the control group, the lower low-density lipoprotein cholesterol by 17.05 % (P < 0.05) and total cholesterol by 9.43 % (P < 0.05) were observed. Additionally, the increase of parameter a* of 13 % (P < 0.05) and the reduction of the natural leakage by 34 % (P < 0.05) were found.

Biomass of Spirulina maxima enriched by biosorption process as a new feed supplement for swine.

This paper deals with the new mineral feed additives with Cu produced in a biosorption process from a semi-technical scale. The natural biomass of edible microalga Spirulina sp. was enriched with Cu(II) and then used as a mineral supplement in feeding experiments on swine to assess its nutrition properties. A total of 24 piglets divided into two groups (control and experimental) were used to determine the bioavailability of a new generation of mineral feed additives based on Spirulina maxima. The control group was feed using traditional inorganic supplements of microelements, while the experimental group was fed with the feed containing the biomass of S. maxima enriched with Cu by biosorption. The apparent absorption was 30 % (P < 0.05) higher in the experimental group. No effect on the production results (average daily feed intake, average daily gain, feed conversion ratio) was detected. It was found that copper concentration in feces in the experimental group was 60 % (P < 0.05) lower than in the control group. The new preparation-a dietary supplement with microelements produced by biosorption based on biomass of microalgae S. maxima-is a promising alternative to currently used inorganic salts as the source of nutritionally important microelements.

Effect of iodine-enriched yeast supplementation of diet on performance of laying hens, egg traits, and egg iodine content.

The aim of the study was to evaluate the effect of iodine yeast (I-yeast) supplementation on the performance, egg traits, and iodine content of eggs of laying hens. The experiment was conducted as a completely randomized design. A total of 60 laying hens (Hy-Line Brown), 25 wk of age, was divided into 3 groups (4 replicates), and a feeding experiment was conducted for 12 wk. The concentrations and forms of iodine added to the basal diet were as follows: control group, 1 mg of iodine/kg of feed, Ca(IO(3))(2)•H(2)O; experimental groups E1 and E2, 1 and 2 mg of iodine per kilogram of feed, I-yeast, respectively. The iodine yeast did not significantly affect BW gain. Lower level of hen day egg production for groups E1 and E2 was not confirmed statistically; however, it was probably the consequence of low replication. Feed intake was the lowest in the E1 group and feed conversion rate was the highest in the E2 group. Furthermore, the egg and albumen weight was the highest in the group supplemented with 2 mg/kg of iodine from I-yeast (P < 0.05). The concentration of iodine in the egg yolk from groups E1 and E2 was respectively about 80 and 90% higher, compared with the control group. Eggshells from the group fed with 2 mg/kg of I-yeast contained almost 3 times more iodine than eggshells from the control group. The results suggest that iodine yeast supplementation in the diet of laying hens is an effective method for increasing iodine concentration in eggs and thus could contribute to elimination of iodine deficiency disorders in humans consuming iodine-enriched eggs.

Effect of macroalgae enriched with microelements on egg quality parameters and mineral content of eggs, eggshell, blood, feathers and droppings.

This study presents results obtained from feeding experiment on laying hens, which were fed with the diet supplemented with two marine macroalgae: Enteromorpha prolifera and Cladophora sp., enriched with microelements [Cu(II), Zn(II), Co(II), Mn(II), Cr(III)]. The applicability of the preparation was tested on five experimental groups of laying hens and one control group. In the control group, microelements were supplemented in the inorganic form, whereas in experimental groups, Cu, Zn, Co, Mn and Cr were replaced by macroalgae enriched with a given microelement ion. During feeding experiment, weight of laying hens, weight of eggs, eggshell thickness and mineral content of blood, feathers, droppings, eggs content (separately yolk and egg white) and eggshell were measured. Also egg number was counted and microclimate (temperature and relative humidity) was monitored. Supplementing bio-metallic feed additives to the diet of laying hens resulted in higher microelement transfer to eggs and enhanced the colour of yolk. It was also found that the presence of Enteromorpha prolifera and Cladophora sp. in laying hens diet influenced advantageously eggs weight, eggshell thickness as well as body weight of hens. On the basis of these results, it could be concluded that Enteromorpha prolifera and Cladophora sp. enriched with microelement ions could be potentially used as mineral feed additives in laying hens feeding.

Biosorption and bioaccumulation of microelements by Riccia fluitans in single and multi-metal system.

In the present study, biosorption and bioaccumulation characteristics of Riccia fluitans to be use as biological mineral feed supplement, was investigated. Preliminary studies showed that R. fluitans was rich in protein (27-31%) and possessed high cation exchange capacity (14.5 mequiv g(-1)) and therefore it has the potential to find an application as biological carrier of microelements that are supplied to feed of animals, the diet of which is deficient in these components. In the present study, various processes of enrichment with microelements of crystalwort were investigated, including biosorption, bioaccumulation by non-growing and growing cells in single-(Cr(III) ions) and multi-metal system (Cr(III), Cu(II), Mn(II), Zn(II) ions). The effect of process parameters (temperature and pH) on metal ions binding efficiency was studied in single-metal system. It was found that at 20 degrees C and pH5 the biomass bound 106 mg g(-1) Cr(III) ions. The experimental results showed that the mostly advantageous process of metal ions binding was biosorption, the process that is also the mostly cost-effective.

The application of ICP-MS and ICP-OES in determination of micronutrients in wood ashes used as soil conditioners.

In the present paper, the elemental composition of wood ashes obtained by the combustion of wood in a fireplace was determined with the use of ICP-MS and ICP-OES techniques. Wood ashes may find a potential application as deacidifying agents and soil conditioners, since they contain calcium (in the form of CaCO(3) and CaO), potassium (in the form of K(2)SO(4) and K(2)CO(3)) and significant levels of micronutrients. However, if applied to soil, it is important to assess the bioavailability of particular elements to plants. This process can be simulated by proper extraction procedures. Various species of wood were combusted in a firestove in a single-family house. The ashes underwent multielemental analyses with ICP-MS Varian Ultra Mass 700 (Australia) and ICP-OES Vista-MPX from Varian (Australia) in order to determine the content of macro- and micronutrients as well as toxic elements. Ashes were also extracted with solutions of 0.1M NaNO(3) and water in order to simulate the process of elemental transfer from ash (used as soil conditioner) to soil solution and consequently to plants. Also, the environmental impact of ash supplementation to soil was assessed in these experiments. Soil was supplemented with 0-20% of ash. After elution, the eluent underwent multielemental analysis by ICP-MS and ICP-OES techniques to determine the content of macronutrients (P, K, Mg), micronutrients (Fe, Mn, Co, Mo, Zn, Cu and Ti) and toxic elements (Hg, Pb, As and Cd). It was shown that fireplace ashes can be applied for deacidification of homestead gardens. Ash may be described as a valuable soil conditioner with N:P:K formula 0:1:3. It is concluded therefore that in order to achieve full fertilization, additional supplementation with nitrogen fertilizer would be necessary.

The application of multielemental analysis in the elaboration of technology of mineral feed additives based on Lemna minor biomass.

Supplementation of microelements (Mn, Zn, Cu, Cr) to livestock diet is of particular concern. There are various mineral feed additives available on the market. The most frequently used are inorganic feed additives, characterized with low bioavailability and high toxicity. Also, organic feed additives are used, including amino acids chelates, in which bioavailability was greatly improved and toxicity reduced. The problem is high price of these products. Therefore, there is the need to search for new biological mineral feed additives with designed composition, that would be characterized with high bioavailability, low toxicity, low cost and that would also possess a nutritional value. Such a possibility offers biological materials. It was found that biomaterials have metal-binding capabilities. Metal ions may be bound to biomass via either biosorption or bioaccumulation process. When elaborating technology of the production of feed additives, containing simultaneously various elements, it is necessary to use the tool of multielemental analysis in order to simultaneously analyze the content of Mn, Zn, Cu, Cr. In the present work, ICP-OES multielemental analysis was used to investigate the process of production of mineral feed additives based on the biomass of aquatic plant Lemna minor. The effectiveness of the processes of biosorption and bioaccumulation of microelements by an aquatic plant L. minor was studied. The mechanism of the process as well as equilibrium was investigated with the use of multielemental analysis by ICP-OES Vista-MPX instrument from Varian (Australia).