Characteristics and phylogenetic analysis of chloroplast genome of a new type of fruit Rubus rosaefolius / 生物工程学报

The genomic DNA of Rubus rosaefolius was extracted and sequenced by Illumina NovaSeq platform to obtain the complete chloroplast genome sequence, and the sequence characteristics and phylogenetic analysis of chloroplast genes were carried out. The results showed that the complete chloroplast genome of the R. rosaefolius was 155 650 bp in length and had a typical tetrad structure, including two reverse repeats (25 748 bp each), a large copy region (85 443 bp) and a small copy region (18 711 bp). A total of 131 genes were identified in the whole genome of R. rosaefolius chloroplast, including 86 protein coding genes, 37 tRNA genes and 8 rRNA genes. The GC content of the whole genome was 36.9%. The genome of R. rosaefolius chloroplast contains 47 scattered repeats and 72 simple sequence repeating (SSR) loci. The codon preference is leucine codon, and the codon at the end of A/U is preferred. Phylogenetic analysis showed that R. rosaefolius had the closest relationship with R. taiwanicola, followed by R. rubraangustifolius and R. glandulosopunctatus. The chloroplast genome characteristics and phylogenetic analysis of R. rosaefolius provide a theoretical basis for its genetic diversity research and chloroplast development and utilization.

Color characterization and its correlation with quality index during ripening of Rubus chingii / 中国中药杂志

The color of Rubus chingii was characterized by digital method, and the content of water extract, alcohol extract, total flavonoids, total polysaccharides, total polyphenols, ellagic acid, linden glycoside, kaophenol-3-O-rutin were determined. Correlation regression was used to analyze the correlation between color and composition. The results showed that L~* was positively correlated with total polyphenols, kaophenol-3-O-rutin and tilide, and moderately positively correlated with total flavones, ellagic acid and aqueous extracts. The a~* value was negatively correlated with total polyphenols, kaophenol-3-O-rutin, and linden glycosides, while was moderately correlated with total flavones, aqueous extracts, and ellagic acid. The b~* value was negatively correlated with the water extract, and moderately correlated with the content of total polyphenols, total polysaccharides, alcohol extract and kaophenol-3-O-rutin, which showed that R. chingii mature color had a significant correlation with material composition in the process of dynamic change. According to the law of dynamic change in the color and quality indexes, it is determined that the appropriate harvest time is in late April to May 1, while the fruit is not turn yellow. The agronomic traits related to fruit was(12.49±0.56) mm in diameter,(14.25±1.19)mm in height,(1.20±0.14) g in weight, the chroma L~* value was 52.87±3.14,a~* value was 2.01±1.58, b~* values was 28.31±3.88. The results lay a foundation for establishing an objective quantitative evaluation model of R. chingii color from experience.

Effective medicinal ingredients and screening of excellent germplasm in Rubus chingii / 中国中药杂志

In order to provide rationale for selection of good germplasm in Rubus chingii, main effective medicinal ingredients of green fruit such as gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside were measured using UPLC for the samples collected from Chun'an county of Zhejiang province, and such parameters as soluble solid contents of ripe fruit of some samples were also measured to study variation among individuals and correlation. It has been found that there were differences among individuals in the contents of gallic acid, ellagic acid, kaempferol-3-rutinoside, astragalin and tiliroside, which ranged from 0.010 2%-0.027 4%, 0.089 5%-0.291 1%, 0.010 5%-0.114 8%, 0.005 8%-0.041 2% and 0.010 9%-0.086 3%, respectively, with a CV of 18.60%, 27.02%, 44.23%, 44.17% and 47.29%, respectively. Gallic acid was positively correlated with ellagic acid, but negatively with kaempferol-3-rutinoside and astragalin significantly. Significantly positive correlation existed between kaempferol-3-rutinoside, astragalin and linden glycoside as well as between ellagic acid and fruit shape index of ripe fruit and between linden glycoside and the content of soluble solids. 51.35% of the individuals had a content of soluble solids more than 15%. Therefore, abundant variations have been found among individuals in effective medicinal ingredients in R. chingii, which shows great potential for selection, but only do 7.61% of the individuals meet the requirement of Chinese pharmacopoeia in terms of the contents of effective medicinal ingredients. Therefore, selection could be first performed in terms of fruit shape index of ripe red fruit, followed by the contents of ellagic acid and kaempferol-3-rutinoside measured. The individuals, in which the contents of effective medicinal ingredients don't meet the requirement of Chinese pharmacopoeia, could be considered for the selection in terms of edible fresh fruit.

Change in malate dehydrogenase and alpha amylase activities in Rubus fruticosus and Valeriana jatamansi treated granary weevil, Sitophilus granarius / Mudança nas atividades de malato desidrogenase e alfa amilase em Rubus fruticosus e gorgulho tratado com Valeriana jatamansi, Sitophilus granarius

Poor storage conditions provide favorable environment to stored grain pests for their growth. The bio-pesticides are the best alternatives to synthetic pesticides. Present study was conducted to compare toxicity of Rubus fruticosus and Valeriana jatamansi against granary weevil, Sitophilus granarius and subsequent changes in enzyme activity responsible for grain damage. In current research 5 g of R. fruticosus fruit and V. jatamansi rhizome powders were tested separately against S. granarius, in 50 g wheat whole grains for seven days in comparison with the control. The enzymatic activity of malate dehydrogenase and α-amylase was observed in the cellular extracts of S. granarius. The insects were crushed and homogenized in phosphate-buffer solution and centrifuged at 10000 rpm for 5 minutes. For the enzymatic measurement supernatant was tested; the spectrophotometer was adjusted at 340 nm. The reagents were mixed and incubated at 25 °C for five minutes. The cuvettes were placed in the experimental and reference sites of spectrophotometer and recorded the change in absorbance for 3-4 minutes. There was 5.60% and 14.92% reduction in the activity of malate dehydrogenase in R. fruticosus and V. jatamansi, treated insects, respectively. The alpha amylase enzyme activity was 6.82% reduced and 63.63% increase in R. fruticosus and V. jatamansi, treated insects, respectively. Present study addresses that both plant powders are effective against granary weevil by altering enzyme activities so both the plant powders can be used as bio-pesticides against the stored grains pests.

As más condições de armazenamento proporcionam um ambiente favorável às pragas armazenadas para o crescimento. Os biopesticidas são as melhores alternativas aos pesticidas sintéticos. O presente estudo foi conduzido para comparar a toxicidade de Rubus fruticosus e Valeriana jatamansi contra gorgulhos, Sitophilus granarius e subsequentes alterações na atividade enzimática responsáveis ​​por danos aos grãos. Na pesquisa atual, 5 g de frutos de R. fruticosus e pós de rizoma de V. jatamansi foram testados separadamente contra S. granarius, em 50 g de grãos integrais de trigo por sete dias, em comparação com o controle. A atividade enzimática da malato desidrogenase e α-amilase foi observada nos extratos celulares de S. granarius. Os insetos foram esmagados e homogeneizados em solução tampão fosfato e centrifugados a 10000 rpm por 5 minutos. Para a medição enzimática, o sobrenadante foi testado; o espectrofotômetro foi ajustado a 340 nm. Os reagentes foram misturados e incubados a 25 °C por cinco minutos. As cubetas foram colocadas nos locais experimentais e de referência do espectrofotômetro e registradas as alterações na absorbância por 3-4 minutos. Houve redução de 5,60% e 14,92% na atividade da malato desidrogenase em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. A atividade da enzima alfa amilase foi reduzida em 6,82% e aumento de 63,63% em R. fruticosus e V. jatamansi, insetos tratados, respectivamente. O presente estudo aborda que ambos os pós de plantas são eficazes contra o gorgulho do celeiro, alterando as atividades enzimáticas, de modo que ambos os pós de plantas possam ser usados ​​como biopesticidas contra pragas de grãos armazenados.

Herbal textual research on Rubi Fructus / 中国中药杂志

Rubi Fructus is a commonly used traditional Chinese medicine. The origin of Rubi Fructus is the dried fruit of Rubus chingii, a plant of the family Rosaceae, according to the 2015 edition of Chinese pharmacopoeia. There are some differences in the plant origin of Rubi Fructus in ancient herbal literature, to trace back its sources, we conducted a textual research on its origin, producing areas, quality evaluation, processing and concocting, properties, tastes and efficacy etc. based on the records of ancient herbal literatures and combined with plant morphology and related investigation. RESULTS:: showed that the variety of Rubi Fructus was more complex among ancient herbal literature, including R. coreanus, R. hirsutus, R. corchorifolius, R. foliolosus and other mixed varieties. Most scholars believe that the R. chingii has not been recorded in ancient herbal literature, while R. chingii was recorded as early as the Ming Dynasty in Compendium of materia medica through our textual research. Ancient Chinese herbs recorded that Rubi Fructus was mostly produced in Hubei, Shandong, Shanxi and Jiangsu provinces, while R. chingii mainly produced in Anhui, Jiangsu, Zhejiang, Jiangxi, Fujian and other provinces nowadays. Also, it was recorded that Rubi Fructus harvested in wheat field during May were the best. Besides, R. chingii with big, full, grain integrate, firm, yellow and green color, sour taste and impurity free possess the best quality in the contemporary. The ancient records of processing and concocting, properties, tastes and efficacy were basically the same as modern ones.These results provide the basis for the correct utilization and further development of Rubi Fructus.

Survival of commercial probiotic strains and their effect on dark chocolate synbiotic snack with raspberry content during the storage and after simulated digestion

BACKGROUND: A key challenge for manufacturers of pro-health food containing active probiotic microorganisms is to develop a product with attractive sensory features along with maintenance of declared number of microorganisms during storage and transfer by alimentary tract. RESULTS: The highest concentration of polyphenols was observed in snacks without an additive of probiotics as well as those with an additive of L. rhamnosus and B. animalis bacteria and concentration of these compounds increased by 9.5% during six months of storage. None of the products distinguished itself in the sensorial assessment although each was assessed positively. The number of microorganisms was stable and comparatively high during six months of storage at a room temperature and in cooling conditions (108 cfu/g). In the digestion model, an influence of aggressive digestion conditions was examined in the alimentary tract on the number of microorganisms, which allowed to arrange strains from the most resistant (S. boulardii) to the most sensitive (B. breve). It must be noted that currently on the market there is no available snack containing probiotic yeast as well as there is no literature data on works on such formulation of food. CONCLUSIONS: In the newly developed snack made of chocolate, in which sugar has been replaced with maltitol, a raw material was added in the form of raspberry, prebiotic in the form of inulin and a strain of probiotic bacteria, including the unprecedented so far S. boulardii, which stands a high chance to occupy a good place on the market of functional food.

Sustancias bioactivas y actividad antioxidante de frutales nativos de páramos y bosques de neblina del norte peruano / Bioactive substances and antioxidant activity of native fruit trees from moorlands and cloud forests in northern Peru

Objetivos. Determinar los principales tipos de sustancias bioactivas, contenido de polifenoles y actividad antioxidante de cinco frutales nativos de paramos y bosques de neblina del norte peruano. Materiales y métodos. Cinco frutales nativos (Gaultheria reticulata «ushpa¼, Hesperomeles obtusifolia «sachón¼, Physalis peruviana «tungay¼, Rubus robustus «muyaca¼ y Symplocos nuda «chimicuno¼") fueron evaluados mediante análisis fitoquímico preliminar, contenido de polifenoles por el método de Folin-Ciocalteau y la actividad antioxidante, mediante tres métodos (DPPH, ABTS y CUPRAC) en tres sistemas de extracción (decocción, etanol 45% y etanol 96% acidulado). Resultados. Los frutales nativos «ushpa¼ y «sachón¼ de los páramos; «muyaca¼ y «chimicuno¼ de los bosques de neblina contienen de 4 a 5 tipos de sustancias bioactivas (compuestos fenólicos, flavonoides, saponinas y triterpenoides/ esteroides). Las especies contienen valores significativamente altos de polifenoles en el extracto acuoso por decocción que se encontraron entre 62,30 ± 2,49 y 5,36 ± 0,20 mg de equivalentes ácido gálico/g de fruto liofilizado. La actividad antioxidante, bajo tres sistemas de extracción y tres métodos de análisis mostró valores significativamente altos entre 1,43 ± 1,19 y 413,92 ± 4,09 mg de equivalentes de Trolox/g fruto liofilizado. Conclusiones. Las sustancias bioactivas identificadas, el contenido de polifenoles y la actividad antioxidante por DPPH, ABTS y CUPRAC, determinan las potencialidades fitoquímicas que le dan valor nutraceútico a estos frutales nativos.

Objectives. To determine the main types of bioactive substances, polyphenol content and antioxidant activity of five native fruit trees of Paramos and Cloud Forests of Northern Peru. Materials and methods. Five native fruit trees (Gaultheria reticulata "ushpa", Hesperomeles obtusifolia "sachón", Physalis peruviana "tungay", Rubus robustus "muyaca" and Symplocos nuda "chimicuno") were evaluated by preliminary phytochemical analysis, polyphenol content by Folin-Ciocalteau method and antioxidant activity by three methods (DPPH, ABTS and CUPRAC) in three extraction systems (decoction, ethanol 45% and ethanol 96% acid). Results. The native fruit trees "ushpa" and "sachon" of the Paramos; "muyaca" and "chimicuno" from Cloud Forests contain 4 to 5 bioactive substances (phenolic compounds, flavonoids, saponins and triterpenoids/steroids). The species contain significantly high values of polyphenols in the aqueous extract by decoction that were found between 62.30 ± 2.49 and 5.36 ± 0.20 mg Gallic Acid equivalents/g of lyophilized fruit. The antioxidant activity, under three extraction systems and three analysis methods, showed significantly high values between 1.43 ± 1.19 and 413.92 ± 4.09 mg of Trolox equivalents/g lyophilized fruit. Conclusions. The bioactive substances identified, the polyphenol content and the antioxidant activity by DPPH, ABTS and CUPRAC, determines the phytochemical potentialities that give nutraceutical value to these native fruit trees.

Rubus sellowii Cham. & Schlitdl. (Rosaceae) fruit nutritional potential characterization / Rubus sellowii Cham. & Schlitdl. (Rosaceae) caracterização do potencial nutricional de frutas

Abstract The aim of this study was to know the physical, chemical and nutritional characteristics of Rubus sellowii (Rosaceae) fruits, known as blackberry, native plant mainly to Rio Grande do Sul State (RS), Brazil. For this, three different populations of this plant were selected and the fruits were analyzed through moisture, ash, titratable acidity, pH, lipids, fibers, carbohydrates, proteins, carotenoids, lycopene, ascorbic acid, aminogram and in vitro digestibility. Fruits showed high acidity (3.28 percent), ash and protein (1.02 and 0.93 percent) and higher ascorbic acid (38.43 mg per 100 g) if compared to Rubus cultivars. Due to Rubus sellowii nutritional properties, they provide benefits to human health.

Resumo O objetivo deste estudo foi conhecer as características físicas, químicas e nutricionais de frutos de Rubus sellowii (Rosáceas), conhecida como amora-preta, planta nativa principalmente do Rio Grande do Sul (RS), Brasil. Para isso, foram selecionadas três populações diferentes desta planta e frutas foram analisadas por meio de umidade, cinza, acidez titulável, pH, lipídios, fibras, carboidratos, proteínas, carotenóides, licopeno, ácido ascórbico, aminograma e digestibilidade in vitro. As frutas apresentaram alta acidez (3,28 g por cento), cinzas e proteínas (1,02 e 0,93 g por cento) e maior ácido ascórbico (38,43 mg por 100 g) em comparação com as cultivares Rubus. Devido às propriedades nutricionais de Rubus sellowii , elas proporcionam benefícios para a saúde humana.

Parâmetros de qualidade em geleias de amora-preta (Rubus sp. ) / Parameters of quality in jams of blackberry (Rubus sp. )

A produção de geleia a partir de frutos de amoreira-preta é uma opção que garante o aproveitamento da matéria prima, que é conhecida por conter elevado valor nutricional. O processamento possibilita o seu consumo durante todos os meses do ano, agregando também um maior valor ao produto final. Foram avaliadas geleias de três marcas comercias com relação aos parâmetros físico-químicos pH, acidez, sólidos solúveis, umidade, sólidos totais, atividade de água e açúcares totais. Dentre as amostras analisadas, observou-se que a amostra C apresentou menor teor de umidade, maior teor de sólidos totais, solúveis, maior grau de doçura e teor de açucares totais. A atividade de água e o potencial hidrogeniônico não apresentaram diferença significativa entre as amostras analisadas.

Processamento e caracterização físico-química de refresco de amora-preta adoçado com mel / Physical-chemical processing and characterization of blackberry refreshment sweetened with honey

Objetivou-se com a presente pesquisa elaborar e caracterizar a composição físico-química de refresco de amora-preta adoçado com mel. Para a elaboração dos refrescos foram utilizados polpa de amora-preta extraída de frutos in natura e mel de abelha Apis Mellifera L., sendo processados em liquidificador industrial e acondicionados em garrafas plásticas de polietileno sob refrigeração. Os refrescos, a polpa e o mel utilizados foram submetidos às análises físico-químicas de acidez total titulável, sólidos solúveis totais, pH, Ratio, atividade de água, cinzas e umidade. Constatou-se que a polpa e mel utilizados na elaboração das bebidas encontravam-se conforme a legislação e foi possível elaborar refrescos de amora-preta com adição de mel, agregando valor as matérias-primas.

Efecto antiespasmódico de Rubus roseus y Mentha pulegium sobre íleon aislado de cobayo (Cavia porcellus) / Antispasmodic effect of Rubus roseus and Mentha pulegium on guinea pig (Cavia porcellus) isolated ileum

Objetivo. El objetivo de este estudio fue investigar el potencial efecto antiespasmódico de Rubus roseus y Mentha pulegium en íleon aislado de cobayo (Cavia porcellus) Materiales y Métodos. Los segmentos de íleon se precontrajeron con acetilcolina (ACh; 4 × 10−6 M) y cloruro de potasio (KCl; 4 × 10-4 M) luego se evaluó la respuesta de los decoctos de Rubus roseus (Rr) Mentha pulegium (Mp) en el baño de órganos aislado en concentraciones crecientes (no acumulativas), como controles positivos se utilizaron nifedipino 5 × 10-8 M (antagonista de canales de Ca2+ voltaje dependientes); atropina sulfato 5,5 × 10-9 M (antagonista muscarínico no selectivo), y N-butilbromuro de hioscina 3 x 10-3 M (antagonista muscarínico M3 selectivo). Se cuantificó la amplitud y la frecuencia de contracción (durante diez minutos) después de la administración de los decoctos. Resultados. Los decoctos de Rubus roseus y Mentha pulegium reducen las contracciones inducidas por acetilcolina (p < 0,001) y KCl (p < 0,01) similares a nifedipino (Mp: 4 y 8 mg/mL); atropina sulfato (Rr: 0,2-4 mg/mL; Mp: 0,8 y 1,6 mg/mL) y N-butilbromuro de hioscina (Mp: 2,4 y 8 mg/mL). Por otro lado, ambos decoctos reducen la frecuencia de contracción inducidas por Ach (p < 0,001) y KCl (p < 0,001). Conclusión. Los decoctos de Rubus roseus y Mentha pulegium tienen efecto antiespasmódico en íleon aislado de cobayo (Cavia porcellus).

Objective. This study aimed to investigate the potential antispasmodic effect of Rubus roseus and Mentha pulegium on isolated guinea pig (Cavia porcellus) ileum. Materials and Methods. The ileum segments were pre-contracted with acetylcholine (ACh, 4 × 10-6 M) and potassium chloride (KCl, 4 × 10-4 M) then the response of the Rubus roseus (Rr) and Mentha pulegium (Mp) decocts was evaluated in the organ bath isolated in increasing concentrations (non-cumulative), as positive controls were used nifedipine 5 × 10-8 M (antagonist of voltage dependent Ca2+ channels) atropine sulfate 5,5 × 10-9 M (non-selective muscarinic antagonist) and 3 x 10-3 M hyoscine N-butylbromide (selective M3 muscarinic antagonist). The amplitude and the frequency of contraction were quantified (during 10 minutes) after the administration of the decocts. Results. The decocts of Rubus roseus and Mentha pulegium decrease the contractions induced by acetylcholine (p < 0.001) and KCl (p <0.01) comparable to nifedipine (Mp: 4 and 8 mg/mL), atropine sulfate (Rr: 0.2 - 4 mg/mL, Mp: 0.8 and 1.6 mg/mL) and hyoscine N-butylbromide (Mp: 2.4 and 8 mg/mL). On the other hand, both decocts reduce the frequency of contraction induced by Ach (p <0.001) and KCl (p <0.001). Conclusions. The decocts of Rubus roseus and Mentha pulegium have an antispasmodic effect on isolated guinea pig ileum (Cavia porcellus).

Protective effect of raspberry extract on ConA-induced acute liver injuryin mice / 中国中药杂志

In this experiment,the antioxidant capacity of raspberry extract and the protective effect on liver injury induced by ConA in mice were investigated. Balb/C male mice were randomly divided into six groups: normal group,model group,bicyclol control group( 200 mg·kg~(-1)),low-dose raspberry extract group( 200 mg·kg~(-1)),middle-dose raspberry extract group( 400 mg·kg~(-1)),and highdose raspberry extract group( 800 mg·kg~(-1)). Each group was intragastrically administered with drugs according to the body weight once a day. Seven days later,all of the groups except for the normal group were treated with ConA( 20 mg·kg~(-1)) through tail vein injection to establish the acute liver injury model. The mice were put to death 8 hours later. The organ indexes were calculated. These rum levels of ALT,AST and LDH and the activities of SOD,CAT,GSH and MDA in liver tissue were detected. HE staining was used to observe the pathological changes of liver tissue in mice. Western blot was used to detect the expressions of Bax,Bcl-2,Nrf2 and Keap-1. The antioxidant capacity of raspberry extract was measured by CAA assay. The results showed that,raspberry extract had a strong antioxidant capacity. Simultaneously,compared with the model group,raspberry extract can significantly improve the pathological conditions of liver,and significantly reduce ALT,AST and LDH activities in serum of liver injury mice( P<0. 01). The activities of SOD,CAT in liver homogenate supernatant were significantly increased in the high-dose group,the content of GSH increased,while the content of MDA was sharply declined in the high-dose group( P<0. 01). Meanwhile,raspberry extract down-regulated the expressions of Bax and Keap-1 and up-regulated the expressions of Bcl-2 and Nrf2. CAA showed that the compound raspberry extract had a strong antioxidant capacity. Therefore,raspberry extract has an obvious protective effect on acute liver injury induced by ConA in mice.

Protein tyrosine phosphatase 1B inhibitory activities of ursane-type triterpenes from Chinese raspberry, fruits of Rubus chingii / 中国天然药物

Protein tyrosine phosphatase 1B (PTP1B) has led to an intense interest in developing its inhibitors as anti-diabetes, anti-obesity and anti-cancer agents. The fruits of Rubus chingii (Chinese raspberry) were used as a kind of dietary traditional Chinese medicine. The methanolic extract of R. chingii fruits exhibited significant PTP1B inhibitory activity. Further bioactivity-guided fractionation resulted in the isolation of three PTP1B inhibitory ursane-type triterpenes: ursolic acid (1), 2-oxopomolic acid (2), and 2α, 19α-dihydroxy-3-oxo-urs-12-en-28-oic acid (3). Kinetics analyses revealed that 1 was a non-competitive PTP1B inhibitor, and 2 and 3 were mixed type PTP1B inhibitors. Compounds 1-3 and structurally related triterpenes (4-8) were further analyzed the structure-activity relationship, and were evaluated the inhibitory selectivity against four homologous protein tyrosine phosphatases (TCPTP, VHR, SHP-1 and SHP-2). Molecular docking simulations were also carried out, and the result indicated that 1, 3-acetoxy-urs-12-ene-28-oic acid (5), and pomolic acid-3β-acetate (6) bound at the allosteric site including α3, α6, and α7 helix of PTP1B.

Antioxidant effects of kimchi supplemented with black raspberry during fermentation protect against liver cirrhosis-induced oxidative stress in rats

BACKGROUND/OBJECTIVES: Oxidative stress is a major effector of various diseases; accordingly, antioxidants are frequently ingested in order to prevent or alleviate disease symptoms. Kimchi contains various natural antioxidants, and it is known that the functional activity varies depending on the ingredients and fermentation state. Black raspberries (BR) contain various bioactive compounds with antioxidant effects. This study investigated the antioxidant and liver-protection effects of kimchi supplemented with black raspberry juice powder (BJP). MATERIALS/METHODS: BJP-added kimchi (BAK; at 0.5%, 1%, and 2% concentrations of BJP) and control (without BJP) were prepared and fermented at 4℃ for 4 weeks. Changes in the antioxidant effects of BAK during fermentation were investigated. In addition, the protective activity of BAK against oxidative stress was investigated in a liver cirrhosis-induced animal model in vivo. RESULTS: BAK groups showed the acidity and pH of optimally ripened (OR) kimchi at 2 weeks of fermentation along with the highest lactic acid bacterial counts. Additionally, BAK groups displayed a higher content of phenolic compounds and elevated antioxidant activities relative to the control, with the highest antioxidant effect observed at 2 weeks of fermentation of OR 1% BAK. After feeding the OR 1% BAK to thioacetamide-induced liver cirrhosis rats, we observed decreased glutamate oxaloacetate transaminase and glutamate pyruvate transaminase activities and elevated superoxide dismutase activity. CONCLUSIONS: These findings showed that the antioxidant effects of OR BAK and feeding of OR 1% BAK resulted in liver-protective effects against oxidative stress.

Estrogenic Activity of Sanguiin H-6 through Activation of Estrogen Receptor α Coactivator-binding Site

A popular approach for the study of estrogen receptor α inhibition is to investigate the protein-protein interaction between the estrogen receptor (ER) and the coactivator surface. In our study, we investigated phytochemicals from Rubus coreanus that were able to disrupt ERα and coactivator interaction with an ERα antagonist. The E-screen assay and molecular docking analysis were performed to evaluate the effects of the estrogenic activity of R. coreanus extract and its constituents on the MCF-7 human breast cancer cell line. At 100 µg/mL, R. coreanus extract significantly stimulated cell proliferation (574.57 ± 8.56%). Sanguiin H6, which was isolated from R. coreanus, demonstrated the strongest affinity for the ERα coactivator-binding site in molecular docking analysis, with a binding energy of

Genotyping-by-sequencing based single nucleotide polymorphisms enabled Kompetitive Allele Specific PCR marker development in mutant Rubus genotypes

Background: Rubus is an economically important fruit crop across the globe. Recently, several Rubus mutant genotypes with improved agronomic traits have been developed using gamma ray irradiation. This study investigated genetic diversity and variations in Rubus mutant genotypes using single nucleotide polymorphism (SNP) markers generated from genotyping-by-sequencing (GBS) analysis. A GBS library of 14 Rubus genotypes, consisting of seven boysenberry mutant lines, four blackberry mutant lines, and three original varieties, were sequenced on the Illumina Hiseq2000 platform. A set of SNPs were analyzed by Kompetitive Allele Specific PCR (KASP) assay in order to discriminate the Rubus genotypes. Results: A total of 50,831,040 (86.4%) reads of clean data were generated, and the trimmed length ranged from 116,380,840 to 509,806,521 bp, with an average of 228,087,333 bp per line. A total of 19,634 high-quality SNPs were detected, which contained 11,328 homozygous SNPs and 8306 heterozygous SNPs. A set of 1504 SNPs was used to perform a phylogenetic analysis, which showed that there were clear differences among the Rubus genotypes based on their origin. A total of 25 SNPs were used for the KASP assays, of which six KASP primer sets were successfully distinguished among the Rubus genotypes. Conclusions: This study demonstrated that the SNP and KASP method is an economically efficient tool for mutant screening in Rubus breeding programs.

Total Saponins of Rubus Parvifolius L. Exhibited Anti-Leukemia Effect in vivo through STAT3 and eIF4E Signaling Pathways / 中国结合医学杂志

OBJECTIVE@#To investigate the anti-leukemia effect of total saponins of Rubus parvifolius L. (TSRP) on K562 cell xenografts in nude mice and the mechanisms of action.@*METHODS@#The K562 cell xenografts in nude mice were established, and then randomly divided into 5 groups, the control group, the cytosine arabinoside group(Ara-c) and 3 TSRP groups (20, 40 and 100 mg/kg). The tumor volume and mass of each group of nude mice were measured and the anti-tumor rates of TSRP were calculated subsequently. The apoptosis status of tumor cells was detected by hematoxylin-eosin (HE) and terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) staining analysis. Finally, the activities of apoptosis related signaling of signal transducer and activator of transcription 3 (STAT3), eukaryotic initiation factor 4E (eIF4E) and B-cell lymphoma-2 (bcl-2) were determined with immunohistochemistry tests.@*RESULTS@#Subcutaneous injection of K562 cells induced tumor formation in nude mice, and the TSRP treated group showed a signifificant inhibitory effect on tumor formation. The nude mice treated with TSRP showed a signifificant decrease in tumor growth rate and tumor weight in comparison to the control group (all P<0.05). The HE staining and TUNEL assay showed that TSRP induced cell death by apoptosis. The immunohistochemical assay showed down-regulation of the bcl-2 gene in the TSRP treated cells. The phosphorylation levels of eIF4E and STAT3 were decreased obviously after the treatment of TSRP.@*CONCLUSION@#TSRP had an excellent tumor-suppressing effect on K562 cells in the nude mice xenograft model, suggesting that TSPR can be developed as a promising anti-chronic myeloide leukemia drug.

Desarrollo y evaluación de una infusión de té rojo (Camellia sinensis) con mora (Rubus ulmifolius) enriquecido con ß-glucanos para el control de glicemia en personas diabéticas / Development and evaluation of an infusion of red tea (Camellia sinensis) with blackberry (Rubus ulmifolius) enriched with ß-glucans for the control of glycemia in diabetic people

La diabetes es una enfermedad crónica degenerativa, exponencialmente creciente, y generando altos costos para los sistemas de salud a nivel mundial. Un alto porcentaje de la población utiliza medicina tradicional como extractos de plantas para satisfacer sus necesidades de atención primaria de salud. El objetivo del estudio fue desarrollar y evaluar una infusión de té rojo con moras enriquecido con ß-glucanos para el control de glicemia en personas diabéticas de la aldea de Jicarito. Se utilizó un DCA con un arreglo factorial 2 × 2 × 2, evaluando dos concentraciones de té rojo con mora (50/50 y 75/25), dos temperaturas (75 y 95 °C) y dos tiempos (2 y 5 minutos), seleccionando el tratamiento con mayor contenido de polifenoles totales (P2T2t2). El mejor tratamiento se evaluó en una muestra de 30 diabéticos, a los cuáles se realizaron medidas antropométricas (peso, talla, circunferencia de cintura-cadera y presión arterial), medidas bioquímicas (glucosa y colesterol) y se analizó la ingesta alimentaria a través de recordatorio de 24 horas. Los resultados demostraron reducción de Índice de Masa Corporal (IMC) en edades de 44- 65 años, en niveles de glucosa en edades de 45-65 y > 65 años, en niveles de colesterol total en edades de 45-65 años y no existió reducción en presión arterial ni en el Índice de Cintura- Cadera (ICC) para ambos sexos. Se determinó una mayor ingesta alimentaria de hidratos de carbono y grasas en la población de diabéticos. Para evaluar modificaciones en medidas antropométricas y bioquímicas se necesita de al menos tres meses de consumo del té.

Efeito de extratos ricos em antocianinas ou elagitaninos de amora silvestre (Morus nigra L. ), amora preta (Rubus spp), e grumixama (Eugenia brasiliensis Lam) no crescimento e na expressão de genes e miRNAs de diferentes linhagens de céluas humanas de câncer de mama / Effects of extracts rich in anthocyanins or ellagitans from mulberry (Morus nigra L. ), blackberry (Rubus spp) and grumixama (Eugenia brasiliensis Lam) on cellular growth and gene and miRNAs expression from distinctive human breast cancer cell lines

O câncer de mama caracteriza-se globalmente como a neoplasia de maior incidência e mortalidade na população feminina. Antocianinas e elagitaninos presentes em frutas como as berries destacam-se por seu promissor efeito protetor em diferentes estágios do desenvolvimento do câncer de mama. Grumixama (G; Eugenia brasiliensis Lam) é uma espécie de cereja nativa do Brasil que assim como as amora-preta (AP; Rubus spp) e silvestre (AS; Morus nigra L.) contém alto teor de antocianinas e elagitaninos. Poucos estudos focaram na ação anticâncer destas berries no câncer de mama. Portanto, o objetivo do presente estudo foi avaliar os efeitos de extratos ricos em antocianinas ou elagitaninos de G, AP e AS no crescimento e na expressão de genes e miRNAs das linhagens de células humanas de câncer de mama MCF-7 (receptor de hormônio positiva) e MDA-MB 231 (receptor de hormônio negativa). Não se observou citotoxicidade após 72 e 96 horas de tratamento com os extratos (25-200µg/mL) ricos em antocianinas (ASANT, APANT e GANT) ou elagitaninos (APELA e GELA), em ambas as linhagens celulares. Após 72 horas de tratamento, GANT e APANT induziram parada de ciclo celular em G0/G1 (12,5 e 50µg/mL, p<0,05) em células MCF-7. Após 96h, ASANT, APANT e GANT induziram parada de ciclo celular em G0/G1 (12,5 e 50µg/mL; p<0,05) nessas mesmas células. Entretanto, na concentração de 200µg/mL apenas GANT induziu parada em G0/G1 (72 e 96h; p<0,05). Em células MDA-MB 231, após 96h APANT e GANT induziram parada de ciclo celular em G0/G1 nas concentrações testadas (12,5, 50 e 200µg/mL, p<0,05), assim como ASANT nas concentrações de 12,5 e 50µg/mL (p<0,05). Em células MCF-7, após 72h APELA e GELA induziram aumento da proporção de células em subG0 (200µg/mL, p<0,05). Em MDA-MB 231, após 72 e 96h, APELA e GELA (200µg/mL) induziram aumento da proporção de células em subG0 (p<0,05) e parada em G0/G1 (p<0,05). Em células MCF-7, GANT induziu morte celular por apoptose (p<0,05) após 72 e 96h de tratamento. Entretanto, em MDA-MB 231 os extratos ricos em antocianinas não induziram morte celular. Em células MCF-7, após 96h GELA e APELA induziram principalmente necrose (p<0,05). Em MDA-MB 231, APELA e GELA induziram apoptose (p<0,05) após 72 e 96h. Em células MDA-MB 231, após 72h de tratamento foi observada inibição da proliferação celular por GELA, GANT e APELA (200µg/mL; p<0,05). Em células MDA-MB 231, 48h de tratamento com GELA; GANT e APELA (200µg/mL) aumentaram a expressão 5 genes (ESR2, FOXA1, JUN, PTGS2,VEGFA) e inibiram a expressão de 10 genes (ADAM23, ATM, BCL2, CDH1, EGF, GLI1, ID1, MKI67, SNAI2 e THBS1) correlacionados ao câncer de mama. Adicionalmente, GELA; GANT e APELA (200µg/mL) induziram aumento da expressão de miR- 210(p<0,05) e APELA (200µg/mL) reduziu a expressão de miRNA 19a/b (p<0,05) em células MDAMB 231. Coletivamente estes resultados sugerem que antocianinas de grumixama e elagitaninos de amora preta e grumixama apresentam potencial efeito protetor contra o câncer de mama. Adicionalmente, essa ação anticarcinogênica pode ser mediada por indução de morte celular, mais especificamente apoptose, redução de proliferação celular e modulação da expressão de genes e miRNAs relacionados ao câncer de mama.

Breast cancer is characterized as the neoplasia with the highest incidence and mortality rates in women worldwide. Anthocyanins and ellagitannins present in certain fruits, such as berries, stand out for their promising protective effect at different stages of breast cancer development. Grumixama (G; Eugenia brasiliensis Lam), a cherry species from Brazil, as well as blackberry (AP; Rubus spp) and mulberry (AS; Morus nigra L.) contain elevated concentrations of anthocyanins and ellagitannins. Few studies focused on the anticarcinogenic action of these berries in breast cancer development. Therefore, the aim of the present study was to evaluate the effects of extracts rich in anthocyanins or ellagitannins from G, AP and AS on cellular growth and genes and miRNAs expression in human MCF-7 (hormone receptor positive) and MDA-MB 231 (hormone receptor negative) cell lines. No cytotoxicity was observed after 72 and 96 hours of treatment with extracts rich in anthocyanin (25-200 µg/mL) (ASANT, APANT and GANT) or ellagitannins (APELA and GELA) in both cell lines. After 72 hours of treatment, GANT and APANT induced cell cycle arrest at G0/G1 (12.5µg/mL and 50µg/mL, p<0.05) in MCF-7 cells. After 96h, ASANT, APANT and GANT induced cycle arrest at G0/G1 (12.5 and 50µg/mL; p<0.05) in MCF-7 cells. However, at 200µg/mL, only GANT induced G0/G1 (72 and 96h; p<0.05). In MDA-MB 231 cells, after 96h APANT and GANT induced cell cycle arrest at G0/G1 with the three tested concentrations (12.5, 50 and 200µg/mL, p<0.05), as well as ASANT at concentrations 12,5 and 50µg/mL (p <0.05). In MCF-7 cells, after 72h APELA and GELA induced an increase in the proportion of cells in subG0 (200µg/mL, p<0.05). In MDA-MB 231, after 72 and 96h, APELA and GELA (200µg/mL) induced an increase in the proportion of cells in subG0 (p<0.05) and cell cycle arrest in G0/G1 (p<0.05). In MCF-7 cells, GANT induced apoptosis (p<0.05) after 72 and 96h of treatment. However, in MDA-MB 231, extracts rich in anthocyanins did not induce cell death. In MCF-7 cells, after 96h GELA and APELA induced mainly necrosis (p<0.05). In MDA-MB 231, APELA and GELA induced apoptosis (p<0.05) after 72 and 96h. In MDA-MB 231 cells, inhibition of cell proliferation by GELA, GANT and APELA (200?g/mL; p<0.05) was observed after 72h of treatment. In MDA-MB 231 cells, treatment for 48h with GELA, GANT and APELA (200µg) increased expression of 5 genes (ESR2, FOXA1, JUN, PTGS2, VEGFA) and inhibited expression of 10 genes (ADAM23, ATM, BCL2, CDH1, EGF, GLI1, ID1, MKI67, SNAI2 and THBS1) correlated with breast cancer. In addition, GELA; GANT and APELA (200µg/mL) induced increased expression of miR-210 (p<0.05) and APELA (200µg/mL) reduced the expression of miRNA 19a/b (p<0.05) in MDA-MB cells 231. Collectively these results suggest that anthocyanins of grumixama and ellagitannins of blackberry and grumixama have potential protective effect against breast cancer. Additionally, this anticarcinogenic action can be mediated by induction of cell death, more specifically apoptosis, reduction of cell proliferation and modulation of the expression of genes and miRNAs related to breast cancer.

Restoration of Declined Immune Responses and Hyperlipidemia by Rubus occidenalis in Diet-Induced Obese Mice

Hyperlipidemia, which is closely associated with a fatty diet and aging, is commonly observed in the western and aged society. Therefore, a novel therapeutic approach for this disease is critical, and an immunological view has been suggested as a novel strategy, because hyperlipidemia is closely associated with inflammation and immune dysfunction. In this study, the effects of an aqueous extract of Rubus occidentalis (RO) in obese mice were investigated using immunological indexes. The mice were fed a high-fat diet (HFD) to induce hyperlipidemia, which was confirmed by biochemical analysis and examination of the mouse physiology. Two different doses of RO and rosuvastatin, a cholesterol synthesis inhibitor used as a control, were orally administered. Disturbances in immune cellularity as well as lymphocyte proliferation and cytokine production were significantly normalized by oral administration of RO, which also decreased the elevated serum tumor necrosis factor (TNF)-α level and total cholesterol. The specific immune-related actions of RO comprised considerable improvement in cytotoxic T cell killing functions and regulation of antibody production to within the normal range. The immunological evidence confirms the significant cholesterol-lowering effect of RO, suggesting its potential as a novel therapeutic agent for hyperlipidemia and associated immune decline.