Journal of Berry Research

Black currants are berries that attract attention due to the beneficial effects they possess. They are proven to be good antioxidants, anticancerogenic, cardio- and neuroprotective agents.

The main objective of this research was to evaluate the differences in the flavonols composition, antioxidant and spasmolytic activity of juices obtained from four different black currant cultivars – Triton, Tenah, Ben Sarek, and Ometa, grown over a 4-year long period on the territory of Serbia.

The qualitative and quantitative determination of black currants juices flavonols was performed using the HPLC. The antioxidative activity was carried out in DPPH and β-carotene systems. The study examined the impact of the main black currants flavonols and black currants juices on the spontaneous, KCl, and acetylcholine-induced ileum smooth muscle contractions.

The dominant flavonol in juices of the four different black currant cultivars was myricetin, with its highest content obtained from the 2015 Triton juice (820.1±34.7 μg g^–1). Quercetin was the second in abundance and the lowest yields were observed in kaempferol.

The investigated black currant cultivars were proven to be good sources of flavonols and potential antioxidants with the significant spasmolytic activity that could be successfully used in treating gastrointestinal disorders.

Monoterpenes and C₁₃-norisoprenoids are key terpenoid compounds for wine aroma. The enzyme encoded by VviDXS1 participates in terpenoid biosynthesis in grapevine fruits and gain-of-function mutations in this gene lead to characteristic muscat aroma.

To assess for VviDXS1 contribution to aroma variation in Northwestern Iberian wine cultivars, we resequenced this gene in 111 cultivars and compared grape juice terpenic composition in 12 of them.

VviDXS1 was capture-targeted for resequencing with Illumina paired-end reads, SAMtools was used for variant calling and gene haplotypes were reconstructed with PHASE. Monoterpenes and C₁₃-norisoprenoids were quantified in free and glycosidically-bound forms from grape juice by gas chromatography-mass spectrometry.

Terpenic composition discriminated between muscat, terpenic and neutral profiles across cultivars. While the terpenic profile of Loureira and Albariño white cultivars was not associated with muscat-like mutations, Albariño carries a V34L substitution in VviDXS1 that is also present in other aromatic cultivars and was not reported before. Tempranillo and Cabernet Sauvignon red cultivars accumulated higher levels of C₁₃-norisoprenoids, which was not associated with specific variation in VviDXS1.

Apart from the uncharacterized substitution present in Albariño, findings suggest that terpenoid pathway-related genes other than VviDXS1 could contribute to the aromatic attributes of these cultivars.

Growing evidence suggests that hyperglycemia could be harmful for cognitive function. That insulin (INS) has a neuro-modulatory role is supported by various findings, but its effect on microglia, the innate immune cells in the brain, is largely unknown. Blueberries have been shown to reduce neuro-inflammation.

We hypothesized that high glucose stimulated an inflammation in microglia and that BB and INS were able to reduce it and both might act through GLUT-1 transporter.

We examined the effects of low (5 mM), medium (25 mM), or high (50 mM) glucose, stimulated or not with lipopolysaccharide (LPS; 100 nM) with either BB extract (2 mg/ml) and/or INS, on inflammatory responses in a microglia cell line. Nitric oxide (NO) production and the expression levels of iNOS, TNF-α, NOX4 and glucose transporter protein-1 (GLUT1) were assessed.

We observed that treatment with BB, similarly to INS treatments, reduced the high glucose concentration-induced response on oxidative stress and inflammation, and that this protective effect is more important with LPS added to glucose media. Interestingly, both BB and INS attenuated the LPS-induced inflammatory response on GLUT1.

Increasing glucose concentration triggers inflammation by microglia. BB as well as INS protected microglia from high glucose levels, by reducing inflammation and altering glucose transport in microglia. These preliminary data compared for the first time BB to Insulin on microglia. Blueberries are promising dietary intervention to prevent diabetic neuropathy. Our preliminary results suggest a possible new mechanism involving GLUT-1 by which BB has insulin-like effects.

HD-Zip genes encode several large and highly conserved protein families of transcription factors that play important roles in plant development and responses to environmental stress. To date, information about the involvement of HD-Zip in grape to drought response is limited.

The production of grapes is limited by a range of biotic and abiotic stresses, which cause significant losses in yield every year as well as a reduction in fruit quality. Identification and analysis of stress related genes in grapes are very important for cultivating more robust varieties with environmental stress resistance

We isolated a homeodomain-leucine zipper gene (HD-Zip), VvHDZ4, from grape, Vitis vinifera L. “Yatomi Rose”, and characterized VvHDZ4 based on analyses of its expression patterns, subcellular localization, transcription activity, and overexpression.

We found that VvHDZ4 was highly expressed in roots and leaves, localized to nucleus, and activated the expression of reporter genes in yeast. The overexpression (OE) of VvHDZ4 led to enhanced drought stress tolerance in Solanum lycopersicum L. “Ailsa Craig” (tomato). The drought-tolerant phenotypes of the OE lines exhibited decreases in electrolyte leakage and rate of water loss, higher photosynthetic production and net photosynthesis rate, and showed up-regulation of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and ascorbate peroxidase (APX), all of which are key antioxidant enzymes known to be activated during drought stress. Moreover, we observed the up-regulation of four additional drought stress indicator genes, SIDREB2A, SIAREB, SIRD29 and SIERD10, in the OE lines. VvHDZ4 improved drought tolerance in tomato, and the modes of action possibly consist of reducing membrane damage, increasing photosynthetic productivity, and modulating the expression of stress defense genes.

This work increases our understanding of the important roles HD-Zip transcription factors in the responses of plants to the environment, especially abiotic stress.

Clostridium difficile Infection (CDI) can lead to diarrhea and fulminant colitis. C. difficile infects the host using toxins. Recent studies report prevalence of CDI in the small intestine. Berries are known to contain antioxidants and phenolic compounds that might mitigate bacterial infection.

We explored the impact of C. difficile toxins on the small intestine using an in vitro approach and used systems biology techniques together with data integration to identify food compounds that can reduce their cytopathic impact.

Differentiated Caco-2 cells were exposed to C. difficile toxins and the transcriptomic changes were studied. To identify foods with potential beneficial counteracting effects, the transcriptomic profiles were integrated with transcriptomics data from Caco-2 cells exposed to various food compounds and analyzed using multivariate analysis.

Beneficial food candidates, selected by multivariate analysis, such as blackcurrant, strawberry and yellow onion were further examined for their potential to counteract the effect of the toxin-induced disruption of cell integrity and toxin translocation. Our results confirmed effects of food compounds, on the cytopathic effects of toxins in the small intestine.

Blackcurrant, strawberry and yellow onion can counteract C. difficile toxins induced effects.

Fruits of Crataegus songarica are commonly used for the treatment of vascular insufficiency and heart problems.

Our aim was to determine the effect of C. songarica on vascular tone and to determine the mechanisms underlying the vasorelaxant properties.

Extracts of C. songarica were tested for vasodilator activity of porcine coronary artery after pre-contraction with the thromboxane mimetic U46619 in the presence or absence of inhibitors of intracellular signaling cascades. Reactive oxygen species were assessed by dihydroethidine staining and the level of eNOS and AKT phosphorylation was measured by immunohistochemical staining.

Extracts of C. songarica berries produced endothelium dependent vasorelaxation, with most significant effect induced by aqueous fraction (AS-CS). This vasorelaxant effect of AS-CS was reduced by inhibition of nitric oxide pathways and inhibition of potassium channels. Inhibition of phosphatidylinositol 3- kinase and Src tyrosine kinase, as well as scavenging of reactive oxygen species, produced an attenuation of the relaxation response. Estrogen receptor antagonists

(tamoxifen and ICI 182,782) reduced the AS-CS mediated vasorelaxation. AS-CS also stimulated the endothelial formation of ROS and phosphorylation of Akt and eNOS.

The data indicated that C. songarica produces an endothelium-dependent vasorelaxation, which is partly dependent upon estrogen receptors, and sensitive to inhibition of ROS/Src/PI3K/NO pathways.

Polyphenols intake increases the function of brown adipose tissue (BAT), stimulating energy expenditure (EE). Calafate (Berberis microphylla) is a polyphenol-rich Chilean native fruit.

To analyse the effect of a treatment with a Calafate extract in the thermogenic activity of mice adipose tissues.

Forty adult C57BL/6J male mice were subdivided into four groups (n=10 each): control diet, control+Calafate (extract: 50mg total polyphenols/kg weight), high-fat diet (HF) and HF+Calafate.

Calafate prevented the increase in body weight and the decrease EE induced by HF. In BAT, Ucp-1 transcript was influenced by the interaction between diet and Calafate (p<0.01), Pparα showed the same expression pattern as Ucp-1 and both, diet (p<0.01) and Calafate (p<0.05), induced significant effects in Sirt1. In inguinal adipose tissue, Pgc1α, Pparα, Prdm16, Sirt1, and Dio2 transcripts presented a decreased expression caused by HF, that was reversed by Calafate. In BAT, an effect of diet (p<0.05) and an interaction between diet and Calafate (p<0.01) was observed in UCP-1 protein levels.

A treatment with Calafate drives less weight gain in mice fed with HF, and reverses the effects generated by it on the expression of thermogenic and browning markers.

Tomatoes are important fruits in the Mediterranean diet and are considered to reduce the risk of different human diseases due to their antioxidative powers.

The purpose of the present study was to investigate the biodiversity within a set of 50 tomato fruit accessions collected across Turkey concerning their antioxidant capacities and the levels of potential health-beneficial compounds including phenolic compounds, carotenoids, ascorbic acid and tocopherols.

All accessions were simultaneously grown in an open experimental field in 2017 and ripe fruits were harvested for analysis. Antioxidant capacities of both hydrophilic and lipophilic extracts were determined using spectrophotometric assays after which individual antioxidants were identified by HPLC using an on-line antioxidant detection system. Phenolic acids, flavonoids, carotenoids and vitamins C and E were quantified using HPLC.

The results indicated that there is a wide diversity within this small collection with respect to their hydrophilic and lipophilic antioxidants.

Hydrophilic antioxidant capacity of the tomatoes was generally related to chlorogenic acid and Vitamin C levels, while lipophilic antioxidants were correlated to all-trans lycopene.

Black raspberry is a vital fruit crop with a high antioxidant function. MADS-box genes play an important role in the regulation of fruit development in angiosperms.

To understand the regulatory role of the MADS-box family, a total of 80 MADS-box genes were identified and analyzed.

The MADS-box genes in the black raspberry genome were analyzed using bioinformatics methods. Through an analysis of the promoter elements, the possible functions of different members of the family were predicted. The spatiotemporal expression patterns of members of the MADS-box family during black raspberry fruit development and ripening were systematically analyzed.

The genes were classified into type I (Mα: 33; Mβ: 6; Mγ: 10) and type II (MIKC^*: 2; MIKC^C: 29) genes. We also obtained a complete overview of the RoMADS-box gene family through phylogenetic, gene structure, conserved motif, and cis element analyses. The relative expression analysis showed different expression patterns, and most RoMADS-box genes were more highly expressed in fruit than in other tissues of black raspberry.

This finding indicates that the MADS-box gene family is involved in the regulation of fruit ripening processes in black raspberry.

Anthocyanins are not only one of the most important antioxidants in plants but also responsible for the coloration in strawberry (Fragaria×ananassa), it is suggested to be associated with ascorbic acid (AsA), sugars and plant hormones metabolism.

To elucidate the molecular differences of genes and pathways in the presence and absence of anthocyanins.

RNA sequencing (RNAseq) of red-fleshed strawberry ‘Benihoppe’ and its white-fleshed mutant ‘Xiaobai’ was performed.

A total of 1,156 and 1,378 transcripts were differentially expressed (DE) in two cultivars at white stage and red stage respectively. Among them, 28, 49 and 72 DE transcripts were involved in AsA, sugars metabolism and plant hormones signaling respectively. Fruits of ‘Benihoppe’ with higher anthocyanins content contained higher AsA, the expression of D-galacturonate reductase (GalUR) in white-fleshed strawberry was down-regulated. Particularly, the expression of transcripts involved in sugars, jasmonic acid (JA) and abscisic acid (ABA) signaling was decreased, by contrast, the expression of transcripts involved in auxin and gibberellins (GAs) signaling was significantly increased.

The lower AsA level in white-fleshed strawberry is possibly because of the lower flux of D-galacturonate biosynthesis pathway. Anthocyanins was associated with gene expression involved in AsA, sugars, and hormones signaling metabolisms.

Increasing evidence has established neuroinflammation as the hallmark of neurodegenerative disorders such as Alzheimer’s disease (AD). However, despite the underlying immunological mechanisms are far from being understood, the involvement of excessive activation of microglia is attracting more and more attention.

In the present study, we investigated the protective effect of black raspberry (BRB) anthocyanins on LPS-induced neuroinflammation in BV2 microglia.

LPS-induced mouse BV2 microglia were treated with black raspberry anthocyanins and the levels of NO, ROS, IL-1β and IL-18 produced by the cells were measured to determine the extent of oxidative stress and inflammatory response.

The results showed that BRB anthocyanins reduced the production of ROS in LPS-induced BV2 microglia by down-regulating the level of NOX2 and its downstream factors, including thioredoxin-interacting protein (TXNIP) and NOD-like receptor protein 3 (NLRP3) inflammasome. Furthermore, BRB anthocyanins inhibited the secretion of Interleukin-18 (IL-18) and Interleukin-1β (IL-1β), eventually attenuating the LPS-induced inflammatory response of BV2 microglia.

BRB anthocyanins might play an important neuroprotective role in inflammation-related neurodegenerative disease, potentially, by down-regulating the NOX2 /TXNIP/ NLRP3 signaling axis in brain microglia.

Red raspberry (Rubus idaeus L.), a natural dietary source of (poly)phenols, has been used as medicine for centuries.

The purpose of this study is to determine the effect of a red raspberry (poly)phenolic extract (RPE) on diet-induced obesity, hepatic steatosis and insulin resistance, and elucidate the underlying molecular mechanisms.

Male specific pathogen-free C57BL/6J mice were randomly divided into three groups (n = 12 per group), and fed with low-fat diet (10% fat energy), high-fat diet (HFD, 45% fat energy), or HFD supplemented with RPE of 150 mg/kg body weight by intragastric administration for 14 weeks. Obesity-related biochemical indexes and hepatic gene expression levels were determined. The statistical analyses were conducted using one-way analysis of variance (ANOVA) followed by Duncan’s multiple range test.

The body weight gain, steatosis grade scores and insulin resistance index in the RPE group decreased by 34.48% (P = 0.00), 58.82% (P = 0.00), and 53.77% (P = 0.00), respectively, compared to those in the HFD group. Moreover, RPE supplement significantly changed the expression profile of the genes involved in lipid metabolism and fibroblast growth factor 21 signaling pathway.

This study demonstrated that RPE protected from diet-induced obesity and related metabolic disorders by improving the lipid metabolism and fibroblast growth factor 21 resistance.